Δ2,3 -1,4 Morpholine-2-carboxylic acid derivatives as antibacterial agents

ABSTRACT

There is described the stereoselective total synthesis of novel Δ 2 ,3 -1,4-morpholine-2-carboxylic acids possessing a fused β-lactam ring in the 1,6-position and carrying a substituent cis to carbon 5 in the 7-position of the fused ring system represented by the general formula ##STR1## wherein Z is halo, hydroxyl, etherified hydroxyl or hydroxyl esterified with a carboxylic acid residue or a sulfonic acid residue and X is amino, azido or acylamino. Also included in the invention are compounds of formula I in which the carboxyl group at the 2-position is protected as by an easily cleavable ester group and salts of both the free acids and carboxyl-protected compounds of formula I. The compounds of formula I are potent antibacterial agents or are of use as intermediates in the preparation of such antibacterial agents.

CROSS-REFERENCE TO RELATED APPLICATION

This is a divisional application of co-pending application Ser. No.567,323 filed Apr. 11, 1975, now U.S. Pat. No. 4,011,216.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The chemical processes of the present invention produce novelantibacterial agents of the β-lactam type containing a hitherto unknownnucleus and useful intermediates for their synthesis.

2. Description of the Prior Art

Penicillins and cephalosporins comprise a group of well-knownantibacterial agents commonly grouped together as a class calledβ-lactam antibiotics. For a recent review of this field with manycitations (especially the first ten) to the prior work, see J. P. Houand J. W. Poole, β-lactam Antibiotics: Their Physiocochemical Propertiesand Biological Activities in Relation to Structure, J. PharmaceuticalSciences, 60(4), 503-532 (April, 1971). Most of the work in this fieldhas fundamentally been done, speaking broadly, with 6-aminopenicillanicacid, 7-aminocephalosporanic acid and derivatives thereof produced byfermentation or chemical transformation of the natural products. Despitethe extensive progress made in preparing active derivatives of6-aminopenicillanic acid and 7-aminocephalosporanic acid, there is acontinuing search for synthetic and semi-synthetic routes to newfamilies of β-lactam antibiotics which may possess more advantageousproperties then those derived from the known penicillin andcephalosporin nuclei.

Considerable work has been done on total chemical synthesis of bothknown β-lactams and nuclear analogs of such known compounds. A recentreview is the text by M. S. Manhas and A. K. Bose, Synthesis ofPenicillin, Cephalosporin C and Analogues, Marcel Decker, Inc., 95Madison Avenue, New York, N.Y., 1969. Another extensive review is by R.B. Morin and B. G. Jackson, Chemistry of Cephalosporin Antibiotics,Fortschr. Chem. Org. Naturst., 28, 343-403 (1970), especially pages379-393; the now famous "Woodward Intermediate" is shown therein ascompound 146 on page 387. A more recent review of β-lactams is that byM. S. Manhas and A. K. Bose, Beta-Lactams: Natural and Synthetic: Part1, Wiley-Interscience, New York, N.Y., 1971. A still further reviewarticle on the synthesis of β-lactams is that by A. K. Kokerjee et al.,Synthesis, 327 (1973).

Other pertinent publications relating to synthesis of β-lactams are:

a. D. M. Brunwin, G. Lowe and J. Parker, J.C.S. Chem. Comm., 1971,865-867, describing synthesis of nuclear analogs of thepenicillin-cephalosporin group.

b. D. M. Brunwin et al., J. Chem. Soc. (C), 1971, 3756-3762 and J.C.S.Chem. Comm., 1972, 589-590 describing total synthesis of nuclear analogsof penicillins and cephalosporins.

c. S. Kukolja, J. Amer. Chem. Soc., 93, 6267-6270 (1971) and 94,7590-7593 (1972) describing preparation of6-phthalimido-5-epipenicillanates and disulfide analogs of penicillins.

d. J. A. Webber et al., J. Medicinal Chemistry, 14(11), 1136-1138 (1971)describing preparation of 3-cyanomethyl cephem nucleus.

e. West German patent specification No. 2,219,601 (Farmdoc 76,051T)describing synthesis of β-lactams of the formula ##STR2## wherein X ishalo, N₃ -- or H₂ N--, A is --S--, --S--CH₂ --, --O--, --O--CH₂ --,--CH₂, --CH₂ CH₂ -- or --NH-- and R¹, R² and R³ are hydrogen, C₁ -C₆alkyl or aryl.

f. U.K. Pat. No. 1,308,822 disclosing β-lactams of the formula ##STR3##where Y = amino or substituted amino.

g. S. Wolfe et al., Can. J. Chem., 50, 2894-2905 (1972) describingsynthesis of sulfur-free penicillin derivatives.

h. French Patent 2,111,859 describing nuclei of the formula ##STR4## and7-acylated derivatives thereof.

i. Helvetica Chimica Acta, 55(2), 388-429 (1972) describing nuclearmodified cephalosporins and penicillins.

j. F. Moll et al., Zeit. fur Naturforsch. B, 27(b)6, 727 (1972)describing nuclear analogs of cephalosporins.

k. U.K. specification Nos. 1,271,013 and 1,271,014 describing γ-lactamsof 7-(acylamino)-3-aminomethyl-ceph-3-em-4-carboxylic acids.

l. U.K. specification No. 1,271,180 describing preparation of novelthiazoline azetidinone rearrangement products useful as intermediates inpenicillin and cephalosporin synthesis.

m. German patent specification Nos. 2,046,822, 2,046,823 and 2,046,824describing synthesis of novel azetidinone intermediates.

n. G. Lowe et al., J. Chem. Soc. Perkins I, 1322 (1973) describing totalsynthesis of nuclear analogs of 7-methylcephalosporins having theformula ##STR5##

o. D. M. Brunwin et al., J. Chem. Soc. Chem. Comm., 865 (1971)describing synthesis of compounds of the formula ##STR6##

p. S. Wolfe et al., Canadian J. Chem., 50, 2902 (1972) describingcompounds of the formula ##STR7##

q. J. P. Luttringer et al., Tetrahedron Letters, 4163-4166 (1973)describing compounds of the formula ##STR8##

r. U.S. Pat. No. 3,835,130 disclosing β-lactams of the formula ##STR9##wherein R_(a) and R_(b) are each hydrogen or R_(a) and R_(b) form acovalent carbon-to-carbon bond, R₁ ' represents hydrogen, R₁ " iscyanoacetyl, bromacetyl or an acyl group of the formula ##STR10## inwhich Ar is phenyl, 3-hydroxyphenyl, 4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 3,5-dichloro-4-hydroxyphenyl or 2-thienyl, R₂' is hydrogen and R₃ ' is hydrogen or lower alkyl.

s. West German Offenlegungsschrift No. 2,355,209 describing synthesis ofβ-lactams of the formula ##STR11## wherein R is acyl, A' is hydrogen,hydroxy, carbamoyloxy, thiocarbamoyloxy, quaternary ammonium, N-loweralkyl carbamoyloxy, N,N-di-lower alkyl carbamoyloxy, N-lower alkylthioand N,N-diloweralkylthiocarbamoyloxy, azido, halo, cyano, a tertiaryamine, acyloxy, or a 5-member heterocyclic thio group having 1-4 heteroatoms; B is H, OCH₃ or SR where R is lower alkyl or aryl; X is adivalent radical selected from --O--, --CH₂ -- or --NY-- where Y ishydrogen, lower alkyl, formyl or benzyl and R' is hydrogen or aprotecting group.

SUMMARY OF THE INVENTION

The present invention provides stereoselective total synthesis ofcertain novel substituted Δ²,3 -1, 4-morpholine-2-carboxylic acidspossessing a fused β-lactam ring in the 1,6-position and carrying asubstituent cis to carbon 5 in the 7-position of the fused ring systemrepresented by the general formula ##STR12## wherein Z is halo,hydroxyl, etherified hydroxyl or hydroxyl esterified with a carboxylicor a sulfonic acid residue and X is azido, amino or acylamino. When X isacylamino, these acids (and their pharmaceutically acceptable salts andphysiologically hydrolyzed esters) are potent antibacterial agents.

Also included in this invention are various novel intermediates usefulin preparing the active β-lactam derivatives described above and variousprocesses for the production of the intermediates and active compounds.

The compounds having the above general formula represent a new family ofβ-lactam antibiotics. They can be considered nuclear analogs ofcephalosporins in which the sulphur atom of the dihydrothiazine ring isreplaced by an oxygen atom and shifted from position 5 to position 4 ofthe β-lactam ring system as numbered in the formula above. Thenomenclature to be used could be the following: ##STR13##

However, Sheehan has used the term O-cephem for the structure ##STR14##[J. C. Sheehan and M. Dadic, J. Heterocyclic Chem., 5, 770 (1968)] andwe propose the use of the term O-2-isocepham for the basic system havingthe formula ##STR15## The numerical prefix indicates the position of thehetero-atom.

To illustrate the above system, the intermediate of the formula##STR16## may be named benzyl 7β-amino-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate and the compound of the formula ##STR17##may be named 7β-(2-aminomethylphenylacetamido)-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylic acid.

There is thus provided by the present invention the novel O-2-isocephemcompounds having the formula ##STR18## wherein R is an acyl group and Zis halo, hydroxyl, hydroxyl esterified with a carboxylic acid or asulfonic acid residue or etherified hydroxyl, and easily cleavableesters and pharmaceutically acceptable salts of said acids and esters.

The acyl group R can be chosen from a wide variety of organic acylradicals which yield products of improved properties and is preferablyan acyl radical which is contained in a naturally occurring orbiosynthetically, semi-synthetically or totally-synthetically producedpharmacologically active N-acyl derivative of 6-aminopenicillanic acidor 7-aminocephalosporanic acid. Examples of suitable acyl groups aredefined in the following general formulae, but it should be noted thatthis is not intended to be an exhaustive list of all the possible acylgroups which may be used.

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

where R^(a) is aryl (carbocyclic or heterocyclic), substituted aryl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl or a nonaromatic or mesoionic heterocyclic group, and n isan integer from 1-4. The preferred R^(a) substituents are (a) arylselected from phenyl, 2-thienyl, 3-thienyl, furyl, 4-isoxazolyl,pyridyl, tetrazolyl, sydnone-3 or -4, imidazolyl, naphthoyl,quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl, thiazolyl,oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl, pyrimidinyl,pyridazinyl or triazinyl; (b) substituted aryl in which the aryl groupsmentioned above under (a) are substituted by one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido,cycloalkylamino, allylamido morpholinocarbonyl, pyrrolidinocarbonyl,piperidinocarbonyl, tetrahydropyridino, furfurylamido orN-alkyl-N-anilino; (c) C₃ -C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂cycloalkyl where the substituents are one or more radicals selected fromchloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁-C₄ alkylamino, C₁ -C₂ alkoxy or amino; (e) C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds; and (f) substituted C₃-C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or 2 double bondsand being substituted by one or more radicals selected from chloro,bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄alkylamino, C₁ -C₂ alkoxy or amino. The most preferred R^(a) groups arephenyl; phenyl substituted by one or more radicals selected from chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy, or trifluoromethyl; 2-thienyl; 3-thienyl; tetrazolyl;sydnone -3; sydnone -4; furyl; isothiazolyl; thiadiazolyl optionallysubstituted with phenyl; oxadiazolyl optionally substituted with phenyl;thiazolyl; imidazolyl; triazolyl; oxazolyl; pyridyl; furazan optionallysubstituted at the 3-position with methoxy; 4-isoxazolyl optionallysubstituted at the 5-position wth methyl and at the 3-position withphenyl or halophenyl; 1,4-cyclohexadienyl; 1-cyclohexenyl and1-aminocyclohexyl.

The most preferred acyl groups of this category are those in which nis 1. Examples of this category include phenylacetyl, halophenylacetyl,nitrophenylacetyl, aminophenylacetyl, β-(o-aminomethylphenyl)propionyl,(lower)alkanoyloxyphenylacetyl (e.g. p-acetoxyphenylacetyl),(lower)alkoxyphenylacetyl (e.g. methoxyphenylacetyl,ethoxyphenylacetyl), (lower)alkylphenylacetyl (e.g. methylphenylacetylor ethylphenylacetyl), hydroxyphenylacetyl (e.g. o-hydroxyphenylacetyl),(lower)alkylaminophenylacetyl (e.g. o-, m- or p-aminomethylphenylacetyl), o- m- or p- guanidinophenylacetyl,o-carboxyphenylacetyl, N,N-bis-(2-chloroethyl)aminophenylpropionyl,thien-2 and 3-ylacetyl, 2- or 3- furylacetyl,1,2,5-thiadiazole-3-acetyl, isothiazolyl-4-acetyl, 4-isoxazolylacetyl,1-cyclohexenylacetyl, 2-aminomethyl-1-cyclohexenylacetyl,1-aminocyclohexylacetyl, 1,4-cyclohexadienylacetyl,2-aminomethyl-1,4-cyclohexadienylacetyl, pyridylacetyl, tetrazolylacetyl(other heterocyclic groups of this type are disclosed in U.S. Pat. Nos.3,819,623 and 3,516,997) or a sydnoneacetyl group as disclosed in U.S.Pat. Nos. 3,681,328, 3,530,123 and 3,563,983. Other groups of this typeinclude 3-phenyl-5-chlorophenyl-5-methylisoxazol-4-ylacetyl and3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylacetyl or a group in whichisoxazolyl is replaced by isothiazole as disclosed in U.S. Pat. No.3,551,440. Still other examples are o-, m- and p-(2'-aminoethoxy)phenylacetyl (as disclosed in U.S. Pat. No. 3,759,905),4,5-dimethoxycarbonyl-1,2,3-triazol-1-ylacetyl or4-cyano-1,2,3-triazol-1-yl-acetyl (as disclosed in U.S. Pat. No.3,821,206) and imidazol-(1)-acetyl (as disclosed in U.S. Pat. No.3,632,810;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

where n is an integer from 1-7. The alkyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom or substituted by, e.g., a cyano group. Examples of this groupinclude cyanoacetyl, valeryl, hexanoyl, heptanoyl, ethoxycarbonyl,octanoyl and butylthioacetyl. A preferred acyl group is cyanoacetyl;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

where n is an integer from 2-7. The alkenyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom. An example of this group is allylthioacetyl; ##STR19## where R^(a)is as defined under (i) and in addition may be benzyl, C₁ -C₆ alkyl or(lower)alkoxycarbonyl and R^(b) and R^(c) which may be the same ordifferent each represent hydrogen, phenyl, benzyl, phenethyl or C₁ -C₆alkyl. The preferred R^(a) substituents in this category are benzyl, C₁-C₆ alkyl, (lower)alkoxycarbonyl and those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups. Themost preferred R^(a) group is phenyl. Examples of this group includephenoxyacetyl, 2-phenoxy-2-phenylacetyl, 2-phenoxypropionyl,2-phenoxybutyryl, benzyloxyacetyl, 2-methyl-2-phenoxypropionyl,p-cresoxyacetyl, p-methylthiophenoxyacetyl and ethoxycarbonylacetyl;##STR20## where R^(a) is as defined under (i) and in addition may bebenzyl or C₁ -C₆ alkyl and R^(b) and R^(c) have the meanings definedunder (iv). The preferred R^(a) substituents in this category arebenzyl, C₁ -C₆ alkyl and those mentioned under (i) as being preferredaryl, substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. The most preferredaryl groups of this type are those in which R^(b) and R^(c) are hydrogenand R^(a) is phenyl; phenyl substituted with one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl; 3-pyridyl; or 4-pyridyl;

    R.sup.a X (CH.sub.2).sub.m CO--                            (vi)

where R^(a) is as defined under (i) and in addition may be benzyl, X isoxygen or sulphur and m is an integer from 2-5. The preferred R^(a)groups are benzyl and those mentioned under (i) as being preferred aryl,substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. An example of thisgroup is S-benzylthiopropionyl.

    R.sup.a CO--                                               (vii)

where R^(a) is as defined under (i). The preferred R^(a) groups arethose mentioned under (i) as being preferred aryl, substituted aryl,cycloalkyl (and substituted cycloalkyl) and cycloalkenyl (andsubstituted cycloalkenyl) groups. The most preferred aryl groups of thiscategory are those in which R^(a) is phenyl; phenyl substituted with oneor more radicals selected from chloro, bromo, iodo, fluoro, nitro,amino, (lower)alkyl, (lower)alkylthio, cyano, (lower)alkoxy,(lower)alkylamino, di(lower)alkylamino, hydroxy, acetoxy ortrifluoromethyl, and most preferably phenyl substituted at the2-position by carboxy or phenyl or at the 2- and 6-positions by methoxy;2-ethoxynaphthoyl; 3-phenyl-5-methylisoxazol-4-yl;3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl and 1-aminocyclohexyl.Examples of this group include 2,6-dimethoxybenzoyl, benzoyl,2-biphenylcarbonyl, 2-aminomethylbenzoyl,2-carboxybenzoyl-2-phenylbenzoyl, 2-thienylcarbonyl, 3-thienylcarbonyland 2-chlorobenzoyl; ##STR21## where R^(a) is as defined under (i) and Yis hydrazino, guanidino, ureido, thioureido and substituted thioureido(as disclosed in U.S. Pat. No. 3,741,962), allophanamido (as describedin U.S. Pat. No. 3,483,188), 3-guanyl-1-ureido (as in U.S. Pat. No.3,796,709) 3-(2-furoyl)ureido, cyanamino (as in U.S. Pat. No.3,796,709), 3-(benzoyl)ureido, azido, amino, acylamino (e.g.carbobenzoxyamino), a group obtained by reacting the amino group of the7-side chain with an aldehyde or ketone (e.g. acetone, formaldehyde,acetaldehyde, butyraldehyde, acetylacetone, methyl acetoacetate,benzaldehyde, salicylaldehyde, methyl ethyl ketone or ethylacetoacetate), hydroxy, etherified hydroxy, esterified hydroxy, carboxy,esterified carboxy (as disclosed for example in U.S. Pat. Nos.3,282,926, 3,819,601 and 3,635,961 and including especially ##STR22##tetrazolyl, cyano, halogeno, acyloxy (e.g. formyloxy or(lower)alkanoyloxy), sulfo, sulfoamino or esterified sulfo. Thepreferred R^(a) substituents are those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred Y substituents are hydrazino; guanidino; ureido; substitutedthioureido of the formula ##STR23## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy- C₁ -C₈ alkyl, (carbo-C₁ -C₈ alkoxy) C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; azido; amino; a group obtained by reacting the aminogroup Y with acetone, formaldehyde, acetaldehyde, butyraldehyde,acetylacetone, methyl acetoacetate, benzaldehyde, salicylaldehyde,methyl ethyl ketone or ethyl acetoacetate; hydroxy; etherified hydroxyincluding especially (lower)alkoxy; carboxy; esterified carboxyincluding especially 5-indanyloxycarbonyl; triazolyl; tetrazolyl; cyano;cyanamino; halogeno; formyloxy; (lower)alkanoyloxy; sulfo; orsulfoamino. Examples of this group include α-aminophenylacetyl;α-carboxyphenylacetyl; 2,2-dimethyl-5-oxo-4-phenyl-1-imidazolyl;α-amino-p-hydroxyphenylacetyl; α-hydroxyphenylacetyl;α-formyloxyphenylacetyl and other aryl groups of this type disclosed inU.S. Pat. Nos. 3,812,116 and 3,821,017; α-amino-α-2- or 3-thienylacetyl;α-amino-α-(3-chloro-4-hydroxy)phenylacetyl;α-amino-α-(1,4-cyclohexadienyl)acetyl; α-azidophenylacetyl;α-amino-α-(1-cyclohexenyl)acetyl; 2-carboxy-α-3-thienylacetyl;α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl; α-amino-α-3- or 4- or5-isothiazolylacetyl (as in U.S. Pat. No. 3,579,506) and other α-aminoand α-hydroxy-heterocyclylacetyl groups as disclosed for example in U.S.Pat. No. 3,821,207; ##STR24## where R^(d), R^(e) and R^(f) which may bethe same or different may each represent C₁ -C₆ alkyl, phenyl orsubstituted phenyl. The preferred phenyl substituents are one or moreradicals selected from chloro, bromo, iodo, fluoro, trifluoromethyl,nitro, amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl,(lower)alkoxyamino, (lower)alkoxy, (lower)alkyl, (lower)alkylamino,hydroxy, (lower)alkylthio, carboxy, di(lower)alkylamino or sulfamyl. Anexample of this group is triphenylmethylcarbonyl. ##STR25## where R^(a)is as defined under (i) and in addition may be hydrogen, C₁ -C₆ alkyl,halogen substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl; benzyl or##STR26## and X is oxygen or sulphur. An example of such a groupCl(CH₂)₂ NHCO; ##STR27## where Y is as defined under (viii) and n is aninteger of 1-4. A most preferred Y substituent is amino. An example ofthis group is 1-aminocyclohexanecarbonyl.

(xii) Aminoacyl, for example

    R.sup.g CH(NH.sub.2)--(CH.sub.2).sub.n CO--

where n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub. 2).sub.m CO--

where m is zero or an integer from 1-10, and n is 0, 1, or 2; R^(g) ishydrogen or an alkyl, aryl, aralkyl or carboxy group or a group asdefined under R^(a) in (i) above; and Ar is an arylene group, e.g.p-phenylene or 1,4-naphthylene. Preferred aryl groups of the aboveformulae are those in which R^(g) is hydrogen, (lower)alkyl, phenyl,benzyl or carboxy and Ar is p-phenylene or 1,4-naphthylene. Examples ofsuch groups are disclosed in U.K. Pat. No. 1,054,806. Examples of groupsof this type include p-aminophenylacetyl and δ-aminoadipoyl derived fromnaturally occurring amino acids and derivatives thereof, e.g.N-benzoyl-δ-aminoadipoyl;

(xiii) Substituted glyoxylyl groups of the formula

    R.sup.h.CO.CO--

where R^(h) is an aliphatic, araliphatic or aromatic group. Thepreferred R^(h) groups are 2-thienyl; 3-thienyl; α-naphthyl;2-phenanthryl or a mono-, di- or tri-substituted phenyl group, thesubstituents being selected from chloro, bromo, iodo, fluoro, amino,di(lower)alkylamino, (lower)alkyl, (lower)alkoxy, nitro or(lower)alkanoylamino. Examples of this category are disclosed in U.S.Pat. Nos. 3,546,219 and 3,573,294. Included in this group are also theα-carbonyl derivatives of the above substituted glyoxylyl groups formedfor example with hydroxylamine, semicarbazide, thiosemicarbazide,isoniazide or hydrazine; ##STR28## where R^(a) has the meaning definedunder (i), X is oxygen or sulphur, X' is oxygen or imino and R^(i)represents (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo(lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenyl of2-6 carbon atoms, ##STR29## n is an integer from 0 to 3 inclusive andeach of R^(k) an R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, bromo, iodo, fluoro or trifluoromethyl. The preferredR^(a) substituents are those mentioned under (i) as being preferredaryl, substituted sryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. Preferred acylgroups of this type are those in which R^(a) is 2-thienyl; 3-thienyl;phenyl; or phenyl substituted by one or more radicals selected fromnitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro or trifluoromethyl; X is oxygen; X' is oxygen orimino and R^(i) is (lower)alkyl, phenyl, 2-thienyl, 3-thienyl, 2-furylor 5-nitro-2-furyl. The most preferred groups are those of the aboveformula where R^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl;X is oxygen; X' is oxygen, and R^(i) is phenyl or 2-furyl. Examples aredisclosed in U.S. Pat. Nos. 3,687,949 and 3,646,024; ##STR30## whereR^(a) has the meaning defined in (i) and R^(i) has the meaning definedin (xiv). The preferred R^(a) substituents are those mentioned under (i)as being preferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred R^(i) substituents include (lower)alkyl, dichloromethyl, C₄-C₇ cycloalkyl, 2-thienyl, 3-thienyl, phenyl, benzyl, halobenzyl,##STR31## Examples of this group are disclosed in U.S. Pat. No.3,626,024 and 3,692,779; ##STR32## where R^(a) has the meaning definedin (i) and R¹ is (lower)alkyl, C₃ -C₁₂ cycloalkyl, aryl (especiallyphenyl), a monocyclic heterocyclic radical having 5 or 6 atoms exclusiveof hydrogen which are C, S, N or O, no more than 2 atoms being otherthan c, or a substituted monocyclic heterocyclic radical as definedabove having one or more substituents selected from halo, (lower)alkyl,(lower)alkoxy or phenyl. Examples of this group are disclosed in U.S.Pat. No. 3,778,436. Most preferred R¹ groups are (lower)alkyl, phenyl,thienyl or furyl.

A preferred class of acyl groups are those of the formula ##STR33##wherein Ar' is a radical of the formula ##STR34## in which R^(m), R^(n)and R^(o) are alike or different and each is hydrogen, hydroxy,(lower)alkyl, cyano, (lower)alkoxy, chloro, bromo, iodo, fluoro,trifluoromethyl, nitro, amino, (lower)alkylamino, di(lower)alkylamino,(lower)alkanoyl, (lower)alkanoyloxy such as p-acetoxy or phenyl and Y isamino or a group obtained by reacting the amino group with acetaldehyde,formaldehyde or acetone, fluoro, chloro, bromo, iodo, hydroxy,(lower)alkanoyloxy, carboxy, guanidino, 3-guanyl-1-ureido,3-(2-furoyl)ureido, 3-benzoylureido, sulfo, sulfoamino, ureido,thioureido, (lower)alkoxy, cyano, cyanamino or indanyloxycarbonyl.Particularly preferred Ar radicals are phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl and1,4-cyclohexadienyl. Particularly preferred Y groups are amino, hydroxyand carboxy. Set forth below are formulae of the most preferred acylgroups of this class: ##STR35## Of most interest are the acyl groups ofthe above class where the acid ArCH(X)COOH is of the D-series.

Other particularly preferred acyl groups for the compounds of formula Iare ##STR36## where U and V are alike or different and each is hydrogen,chloro or fluoro; ##STR37##

Substituent Z in formulae I and II above may be halo (chloro, bromo,fluoro or iodo), hydroxyl, hydroxyl esterified with a carboxylic acid ora sulfonic acid residue or etherified hydroxyl. Esterified hydroxylgroups include radicals of the formula

    --OR.sub.1

wherein R₁ is a acyl group, a (lower)alkylsulfonyl group, anarylsulfonyl group or an aralkylsufonyl group and are preferably thoseof the formula

    --OCOR.sub.2 or --OSO.sub.2 R.sub.2 '

wherein R₂ is hydrogen, amino, (lower)alkyl (e.g. methyl, ethyl, propyl,isopropyl, butyl, isobutyl, t-butyl, amyl, hexyl), C₃ -C₇ cycloalkyl(e.g. cyclobutyl, cyclopentyl, cyclohexyl, cyclopropyl or cycloheptyl),C₃ -C₇ cycloalkyl(lower)alkyl (e.g. cyclobutylmethyl, cyclobutylethyl,cyclopentylmethyl, cyclohexylmethyl, cyclopentylpropyl, etc.), aryl(e.g. phenyl or naphthyl), aralkyl (e.g. benzyl, tetrazolylacetyl,2-(1-naphthyl)ethyl or phenethyl), or aryloxyalkylene, e.g.,phenoxymethyl, and R₂ ' is (lower)alkyl, aryl or aralkyl. The R₂ and R₂' groups above may be optionally substituted as by one or more(lower)alkoxy, (lower)alkylthio, halogen, (lower)alkyl, nitro, hydroxy,acyloxy, carboxy, amino, (lower)alkylamino or acylamino radicals. Themost preferred R₂ groups are hydrogen, (lower)alkyl (especially methyl)and amino. The most preferred R₂ ' groups are (lower)alkyl (especiallymethyl), trifluoromethyl and p-tolyl.

Etherified hydroxyl groups include radicals of the formula

    --OR.sub.3

where R₃ is (lower)alkyl, (lower)cycloalkyl,(lower)cycloalkyl-(lower)alkyl, aryl, aralkyl or a heterocyclic group,any of said R₃ groups bring optionally substituted by one or more(lower)alkoxy, (lower)alkoxy(lower)alkyl, (lower)alkylthio, halogen,(lower)alkyl, (lower)cycloalkyl, (lower)alkenyl, nitro, hydroxy,acyloxy, carboxy, amino, di(lower)alkylamino, (lower)alkylamino,trifluoromethyl, aryl, aralkyl or acylamino radicals.

Preferred R₃ groups include benzyl and 5- or 6-membered heterocyclicradicals containing 1-4 atoms selected from N, O and S, saidheterocyclic radicals being optionally substituted by one or moresubstituents selected from halogen, amino, nitro, C₁ -C₄ alkyl, C₃ -C₄cycloalkyl, C₁ -C₄ alkoxy, C₂ -C₄ alkenyl, trifluoromethyl phenyl,benzyl, C₁ -C₄ alkylthio, C₁ -C₄ alkylamino, di C₁ -C₄ alkylamino oralkoxyalkyl of up to 4 carbon atoms. Examples of preferred heterocyclicR₃ groups include furyl, thienyl, pyrazolyl, imidazolyl, isoimidazolyl,triazolyl, tetrazolyl, thiazolyl, thiadiazolyl, thiatriazolyl, oxazolyl,oxadiazolyl, isothiazolyl, isoxazolyl, pyridyl, pyridazinyl, pyrazinyl,pyrimidinyl and triazinyl, said radicals being optionally substituted byone or two or the above-mentioned substitutents, most preferably C₁ -C₄alkyl, C₁ -C₄ alkoxy or trifluoromethyl groups. Particularly preferredheterocyclic R₃ groups include 1,2,3-triazolyl, 1,2,4-triazolyl,1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,3,4-oxadiazolyl,1,2,3,4-tetrazolyl, 2-methyl-1,3,4-thiadiazol-5-yl,2-methyl-1,3,4-oxadiazol-5-yl and 1-N-methyltetrazolyl.

Preferred Z substituents in formula I and II are those of the formulae-halo, i.e. chloro, bromo, iodo, fluoro; --OH; --OCHO; --OCOCH₃ ; --OSO₂CH₃ ; --OSO₂ CF₃ ; --OSO₂ C₆ H₄ CH₃ (para); --OCH₂ C₆ H₅ ; and --OCONH₂.

When Z is hydroxyl, compounds of formulae I and II may also exist as thelactones which are formed by internal esterification with the carboxylgroups.

The term "(lower)alkyl" as used herein means both straight and branchedchain aliphatic hydrocarbon radicals having from one to ten carbon atomssuch as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,amyl, hexyl, 2-ethylhexyl, heptyl, decyl, etc. Similarly, where the term"(lower)" is used as part of the description of another group, e.g."(lower)alkoxy", it refers to the alkyl portion of such group which istherefore described above in connection with "(lower)alkyl".

The pharmaceutically acceptable salts referred to above include thenontoxic carboxylic acid salts, e.g. nontoxic metallic salts such assodium, potassium, calcoum and aluminum, the ammonium salt and saltswith nontoxic amines, e.g. trialkylamines, procaine, dibenzylamine,N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylenediamine,N-alkylpiperidine and other amines which have been used to form salts ofpenicillins and cephalosporins. When a basic group is present, as whenit occurs in the 7-acyl group, the present invention also includes thepharmaceutically acceptable acid addition salts, e.g. salts with mineralacids such as hydrochloric, hydrobromic, hydroiodic, phosphoric,sulfuric and salts with organic acids such as maleic, acetic, citric,oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic andmalic. The term "pharmaceutically acceptable salts" is also meant toinclude nontoxic acid addition salts of the easily cleavable estersreferred to above. The compounds which contain a basic group in radicalR may also be present in the form of an internal salt, i.e. in the formof the zwitterion.

The easily cleavable esters referred to above include ester groups whichare removable by methods, e.g. chemical or enzymatic hydrolysis,treatment with chemical reducing agents under mild conditions,irradiation with ultraviolet light or catalytic hydrogenation, which donot result in any appreciable destruction of the remaining portion ofthe molecule. Examples of suitable esters include those disclosed inU.S. Pat. Nos. 3,284,451 and 3,249,622 and U.K. Pat. Nos. 1,229,453 and1,073,530. Esters which have been used previously in penicillin andcephalosporin chemistry include for example benzhydryl, p-nitrobenzyl,benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl,p-methoxybenzyl, acetonyl, phthalidyl, indanyl and (lower)alkyl such asmethyl, ethyl and t-butyl. Particularly preferred easily cleavableesters are those which are hydrolyzed under physiological conditionssuch as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl andmethoxymethyl.

As the O-2-isocephem compounds of the present invention may possess oneor more asymmetric carbon atoms, the invention includes all of thepossible enantiomeric and diastereomeric forms of the compounds of thegeneral formula II shown above. Resulting mixtures of isomers can beseparated into the individual isomers according to methods which areknown per se, e.g. fractional crystallization, adsorption chromatographyor other suitable separation processes. Resulting racemates can beseparated into the antipodes in the usual manner after introduction ofsuitable salt-forming groupings, e.g. by forming a mixture ofdiastereoisomeric salts with optically active salt-forming agents,separating the mixture into diastereoisomeric salts, and converting theseparated salts into the free compounds, or by fractionalcrystallization from optically active solvents.

It will be appreciated that certain of the compounds of this inventionexist in various states of solvation and the anhydrous as well assolvated forms are within the scope of the invention.

The free acid compounds of the above general formula II where R is acyland physiologically hydrolyzed esters thereof together with thepharmaceutically acceptable salts of such free acids and esters areuseful as antibacterial agents. Certain of the 7-acylated compounds offormula II, e.g. those in which Z is halo, hydroxyl, --OCHO, --OCH₂ C₆H₅ or sulfonyloxy (especially --OSO₂ CH₃, --OSO₂ CF₃ or --OC₆ H₄ CH₃)are useful not only as active antibacterial agents per se but asintermediates in preparation of other active 7-acylated derivativeswithin the scope of formula II. The remaining compounds of the abovegeneral formula II including salts and esters thereof are valuableintermediates which can be converted into the above-mentionedpharmacologically active compounds in a simple manner for example, asdescribed below.

Preferred embodiments of the present invention are the compounds of theformula ##STR38## wherein R is an acyl group and R₄ is halo, hydroxy,--OCHO, --OCONH₂, --OCOCH₃, --OCH₂ C₆ H₅ or --OSO₂ CH₃, and easilycleavable esters and pharmaceutically acceptable salts thereof.

Preferred compounds of formula IIa are those in which R is an acyl groupselected from the acyl groups defined above under (i) ot (xvi). Use ofthe acyl groups mentioned above as being preferred within categories (i)or (xvi) results in active end-products having the most advantageouspharmacological properties.

More preferred acids, esters and salts of formula IIa are those in whichacyl group R is ##STR39## in which Ar' is a radical of the formula##STR40## in which r^(m), R^(n) and R^(o) are alike or different andeach is hydrogen, hydroxy, (lower)alkyl, cyano, (lower)alkoxy, chloro,bromo, iodo, fluoro, trifluoromethyl, nitro, amino, (lower)alkylamino,di(lower)alkylamino, (lower)alkanoyl, (lower)alkanoyloxy such asp-acetoxy or phenyl and Y is amino or a group obtained by reacting theamino group with acetaldehyde, formaldehyde or acetone; fluoro; chloro;bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy; guanidino;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido; sulfo;sulfoamino; ureido; thiouriedo; (lower)alkoxy; cyano; cyanamino; orindanyloxycarbonyl.

Other preferred acids, esters and salts of formula IIa are those inwhich R is ##STR41## wherein Ar' is phenyl, p-hydroxyphenyl,4-hydroxy-3-, 5-dichlorophenyl, o-, m- or p- aminomethylphenyl,2-thienyl, 3-thienyl, 1-cyclohexenyl or 1,4-cyclohexadienyl and Y isamino, carboxy or hydroxy.

Other preferred compounds of formula IIa are those wherein R is an acylgroup of the formula ##STR42##

Other preferred compounds of formula IIa are those wherein R is an acylgroup of the formula ##STR43## wherein U and V are alike or differentand each is hydrogen, chloro or fluoro; ##STR44##

A most preferred group of compounds are those acids defined by formulaIIa wherein R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-theinyl(acetyl, α-amino-α-(3-theinyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-theinyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.

Another most preferred group of compounds are the D-isomers of thoseacids defined by formula IIa wherein R isα-amino-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxyα-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.

Preferred compounds of formula IIa are the acids in which R isphenoxyacetyl, or the pharmaceutically acceptable salts thereof.

Other preferred compounds of formula IIa are the acids in which R isphenylacetyl, or pharmaceutically acceptable salts thereof.

Other preferred compounds of formula IIa are the acids in which R is2-thienylacetyl or 3-thienylacetyl, or pharmaceutically acceptable saltsthereof.

Still further preferred compounds of formula IIa are the acids in whichR is α-hydroxyphenylacetyl or α-aminophenylacetyl, or pharmaceuticallyacceptable salts thereof. The pivaloyloxymethyl, phthalidyl, indanyl,acetoxymethyl and methoxymethyl esters of these acids as well aspharmaceutically acceptable salts thereof are also preferred compoundsof the present invention. The isomers of those compounds in which theα-carbon atom of the 7-acyl group is of the D-series are found to be ofparticular importance.

The present invention further provides various novel intermediatesuseful in the synthesis of the 7-acylamido O-2isocephem compounds offormula II described above.

Preferred embodiments of the present invention are the novelintermediates having the formula ##STR45## wherein Z is halo, hydroxyl,hydroxyl esterified with a carboxylic acid or a sulfonic acid residue oretherified hydroxyl and R" is hydrogen or an easily cleavable estercarboxyl-protecting group, and salts thereof. The preferred Zsubstituents are as defined above in connection with the compounds offormula II.

The most preferred intermediates of formula IV are those of the formula##STR46## wherein R₄ is halo, hydroxyl, --OCHO, --OCH₂ C₆ H₅, --OCONH₂,--OCOCH₃ or --OSO₂ CH₃.

Intermediates of formula IVa in which R₄ is --OSO₂ CH₃ or --OCHO are ofparticular importance as starting materials in preparing otherintermediates within the scope of formula IV.

Other preferred intermediates are the compounds having the formula##STR47## wherein Z is halo, hydroxyl, hydroxyl esterified with acarboxylic acid or a sulfonic acid residue or etherified hydroxyl and R"is hydrogen or an easily cleavable ester carboxyl-protecting group, andsalts thereof. The preferred Z substituents are those mentioned above asbeing preferred in connection with the compounds of formula II.

The most preferred intermediates of formula III are those of the formula##STR48## wherein R₄ is halo, hydroxyl, --OCHO, --OCH₂ C₆ H₅, --OCOCH₃,--OCONH₂ or --OSO₂ CH₃ and R is hydrogen or an easily cleavable estercarboxyl-protecting group.

Preferred intermediates of formula IIIa are those in which R₄ is --OCHOor --OSO₂ CH₃.

The intermediates of formulae III and IV may be in the form of the freecarboxylic acid or a salt thereof or in the form where the carboxylgroup is protected in a conventional manner such as preferably byesterification. The protecting group is selected so that it may beremoved by methods which do not result in any appreciable destruction ofthe remaining portion of the molecule. Preferred carboxyl protectinggroups are the easily cleavable esters as defined above including inparticular benzhydryl, p-nitrobenzyl, trichloroethyl, silyl includingespecially trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl,(lower)-alkyl such as methyl, t-butyl or ethyl, benzyl, triphenylmethyl,methoxymethyl, acetoxymethyl, phthalidyl, indanyl and pivaloyloxymethyl.

The novel 7-acylamido compounds of formula II may be prepared byN-acylating a 7-amino intermediate of the formula ##STR49## wherein Z ishydroxyl, hydroxyl esterified with a carboxylic acid or a sulfonic acidresidue or etherified hydroxyl and R" is hydrogen or an easily cleavableester carboxyl-protecting group, or a salt thereof, with an acylatingacid of the formula

    R--COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amine and, if desired, converting theso-produced product to the corresponding 7-acylated product having asubstituent Z different from that in starting material III and, ifdesired, (a) when R" is a carboxyl-protecting group, converting the7-acylated ester to the free acid compound or a physiologicallyhydrolyzed ester or a pharmaceutically acceptable salt of said acid orester, or (b) when R" is hydrogen, converting the 7-acylated carboxylicacid to a physiologically hydrolyzed ester or a pharmaceuticallyacceptable salt of said acid or ester and, if desired, resolving aresulting isomer mixture into its component isomers.

The 7-amino starting materials of general formula III are of useprimarily as intermediates in preparing the pharmacoligically activeN-acyl derivatives of formula II. The free acids, physiologicallyhydrolyzed esters and pharmaceutically acceptable salts of said acidsand esters of formula III, however, do possess some antibacterialactivity per se against various pathogenic microorganisms.

The 7-acylamido O-2-isocephem compounds of formula II are prepared byN-acylation according to known methods of the 7-amino group ofintermediate III with an acylating acid of the formula

    R--COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amino group. The acylating agents forpreparing the products of formula II are known, readily preparable byknown methods or described herein.

Intermediate III may be acylated either in the form of the freecarboxylic acid (or salt thereof) or as an easily cleavable ester (oracid addition salt thereof). Preferred esters include benzhydryl,benzyl, p-nitrobenzyl, trichloroethyl, silyl (especiallytrimethylsilyl), phenacyl, p-methoxybenzyl, acetonyl, (lower)alkylincluding particularly methyl, ethyl and t-butyl, triphenylmethyl,methoxymethyl, acetoxymethyl, pivaloyloxymethyl, phthalidyl and indanyl.The procedures for preparing esters of carboxylic acids are disclosed inthe literature and are well-known to those skilled in the art ofpenicillin and cephalosporin chemistry. Methods for preparing certain ofthe more preferred easily cleavable esters, i.e. the pivaloyloxymethyl,acetoxymethyl, methoxymethyl, acetonyl and phenacyl esters, aredisclosed in U.S. Pat. No. 3,284,451 and in U.K. Pat. No. 1,229,453.Preparation of phthalidyl esters of penicillins and cephalosporins isdescribed in South African patent application Nos. 72/3799 and 72/3800.The free acid form of intermediate III may also be converted to a silylester, e.g. trimethylsilyl ester, as by the methods described in theliterature, e.g. U.S. Pat. No. 3,249,622. The silyl estercarboxyl-protecting group may be easily removed following the acylationreaction by hydrolysis or alcoholysis.

Prior to the acylation reaction, any reactive substituents on theacylating acid or derivative thereof, e.g. hydroxyl, carboxyl ormercapto, may be protected by use of suitable protecting or blockinggroups which are well-known to those skilled in the art of β-lactamchemistry, e.g. as by acylation or silylation. When the acylating agentcontains an amino functional group in the acyl moiety, the amino groupis protected by a conventional amino-blocking group which may be readilyremoved at the conclusion of the reaction. Examples of suitableamino-protecting or blocking groups include t-butoxycarbonyl,carbobenzyloxy, 2-hydroxy-1-naphthcarbonyl, trichloroethoxycarbonyl,2-ethoxycarbonyl-1-methylvinyl and 2-methoxycarbonyl-1-methylvinyl. Aparticularly valuable amino-blocking group is a proton, as in theacylating agent of the formula ##STR50## Preferred amino-protectinggroups are t-butoxycarbonyl, carbobenzyloxy, the proton and a β-diketoneor β-ketoester as in U.K. Pat. No. 1,123,333 or U.S. Pat. Nos. 3,325,479and 3,316,247, e.g. methyl acetoacetate, or a β-ketoamide as in JapanPat. No. 71/24714. When the t-butoxycarbonyl, carbobenzyloxy,β-ketoester, β-diketone or β-ketoamide protecting groups are employed,it is preferred to convert the acylating acid containing the blockedamino group to a mixed anhydride, e.g. with ethyl or isobutylchloroformate, before reaction with compound III or a salt thereof.After the acylation coupling reaction, the amino-protecting group andany other functional protecting groups used may be removed by methodsknown per se to form the desired product of formula II. With respect toamino-protecting groups, the t-butoxycarbonyl group may be removed byuse of formic acid, the carbobenzyloxy group by catalytic hydrogenation,the 2-hydroxy-1-naphthcarbonyl group by acid hydrolysis, thetrichloroethoxycarbonyl group by treatment with zinc dust in glacialacetic acid, the proton by neutralization, etc.

Acylation of a free amino group of a cephalosporin or penicillin nucleusis a well-known reaction, and any of the functional equivalents of thecarboxylic acid RCOOH commonly used in penicillin or cephalosporinchemistry as acylating agents for primary amino groups may be employedin acylating intermediate III. Examples of suitable acylatingderivatives of the free acid include the corresponding acid anhydrides,mixed anhydrides (e.g. alkoxyformic anhydrides), acid halides, acidazides, active esters and active thioesters. The free acid may becoupled with compound III after first reacting said free acid withN,N'-dimethylchloroformininium chloride [cf. Great Britain Pat. No.1,008,170 and Novak and Weichet, Experientia XXI, 6, 360(1965)] or bythe use of enzymes or of an N,N'-carbonyldiimidazole or anN,N'-carbonylditriazole [cf. South African Specification 63/2684] or acarbodiimide reagent [especially N,N'-dicyclohexylcarbodiimide,N,N'-diisopropylcarbodiimide orN-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide: cf. Sheehan and Hess,J.A.C.S., 77, 1967 (1955)], or of alkylylamine reagent [cf. R. Buijleand H. G. Viehe, Angew. Chem. International Edition 3, 582, (1964)] orof an isoxasolium salt reagent [cf. R. B. Woodward, R. A. Olofson and H.Mayer, J. Amer. Chem. Soc., 83, 1010 (1961)], or of a ketenimine reagent[cf. C. L. Stevens and M. E. Munk, J. Amer. Chem. Soc., 80, 4065 (1958)]or of hexachlorocyclotriphosphatriazine orhexabromocyclotriphosphatriazine (U.S. Pat. No. 3,651,050) or ofdiphenylphosphoryl azide [DPPA; J. Amer. Chem. Soc., 94, 6203-6205(1972)] or of diethylphosphoryl cyanide [DEPC; Tetrahedron Letters No.18, pp. 1595-1598)] or of diphenyl phosphite [Tetrahedron Letters No.49, pp. 5047-5050 (1972)]. Other examples of suitable amide couplingreagents which have been described in the literature include (CH₃)₂ SCH₂CCHBr/DMSO (J. Chem. Soc. (C) 1904 (1969), HCCOCH₃ (Rec. Trav. Chim. 74,769 (1955), (CH₃)₂ C(OCH₃)₂ (Chim. Ther. 2, 195 (1967), SiCl₄ (J. Org.Chem. 34, 2766 (1969), TiCl₄ (Can. J. Chem. 48, 983 (1970), (PNCl₂)₃ (J.Org. Chem. 33, 2979 (1968), SO₃.DMF (J. Org. Chem. 24, 368 (1959), ionexchange resins (Helv. 44, 1546 (1961) and J.C.S. C, 874 (1969) and##STR51## (J. Chem. Soc. 4650 (1964). An equivalent of the acid chlorideis the corresponding azolide, i.e., an amide of the corresponding acidwhose amide nitrogen is a member of a quasi-aromatic five membered ringcontaining at least two nitrogen atoms, i.e., imidazole, pyrazole, thetriazoles, benzimidazole, benzotriazole and their substitutedderivatives. As an example of the general method for the preparation ofan azolide, N,N'-carbonyldiimidazole is reacted with a carboxylic acidin equimolar proportions at room temperature in tetrahydrofuran,chloroform, dimethylformamide or a similar inert solvent to form thecarboxylic acid imidazolide in practically quantitative yield withliberation of carbon dioxide and one mole of imidazole. Dicarboxylicacids yield dimidazole. The by-product, imidazole, precipitates and maybe separated and the imidazolide isolated, but this is not essential. Apreferred acylating agent for preparing 7-acylamido compounds containingan α-amino substituent, e.g. α-aminobenzyl, α-amino-α-thienylmethyl,etc. is the N-carboxy anhydride (Leuch's anhydride). In this structurethe group which activates the carboxyl group also serves to protect theamino group. Another preferred acylating agent for introducing a sidechain containing an α-amino functional group is the acid chloridehydrochloride, of the formula ##STR52## which also serves a dualfunction of carboxyl activation and amino protection. Mention was madeabove of the use of enzymes to couple the free acid with compound III.Included in the scope of such processes are the use of an ester, e.g.the methyl ester, of that free acid with enzymes provided by variousmicroorganisms, e.g. those described by T. Takahashi et al., J.A.C.S.,94(11), 4035-4037 (1972) and by T. Nara et al., J. Antibiotics (Japan)24(5), 321-323 (1971) and in U.S. Pat. No. 3,682,777. A particularlypreferred coupling agent for coupling the acylating acid with compoundIII (or a salt or ester thereof) isN-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) as described inJ.A.C.S., 90, 823-824 and 1652-1653 (1968) and U.S. Pat. No. 3,455,929.

The particular process conditions, e.g. temperature, solvent, reactiontime, etc. selected for the coupling reaction are determined by thenature of the reactants and acylation method used and are known to thoseskilled in the art.

The acylating agents which may be used to form the N-acyl compounds offormula II are known in the literature along with methods for theirsynthesis or are disclosed in the examples which follow. In those caseswhere the acylating agent contains one or more asymmetric carbon atomsand thus exists in optically active forms, the compounds obtained usingsuch an acylating agent are ordinarily obtained in racemic form. Whenthe separate optical isomers are desired, the acylating agent can beresolved in a conventional manner such as by reacting the free acid withcinchonine, strychnine, brucine or the like, fractionally crystallizingto separate the diastereoisomeric salts and separately acidifying thesolid phase and the liquid phase to liberate the optical isomers.

The 7-acylamido compounds of the present invention may be isolated inany of the ways customarily employed for the isolation of correspondingcephalosporin compounds. Formation of a desired pharmaceuticallyacceptable carboxylic acid or acid addition salt is carried out by knownmethods, e.g. reaction of the acid of compound II (or ester in the caseof acid addition salts) with an appropriate base or acid.

At the conclusion of the acylation reaction, the product obtained may beconverted (before or after removal of any protecting groups) by methodsknown per se to another desired product of formula II.

A compound of the formula ##STR53## in which Z is hydroxyl and R' is aneasily cleavable carboxyl-protecting group may be converted by acylationto the corresponding 7-acylamido compound in which Z is hydroxylesterified with a carboxylic acid or a sulfonic acid residue. Acylationof the 3-hydroxymethyl group to produce carboxylic esters is preferablycarried out using an acid anhydride, e.g. acetic anhydride, in thepresence of an organic base such as pyridine. Other conventionalacylating agents may be used including acid halides (preferably acidchlorides), mixed anhydrides or free acids in the presence of condensingagents. Methods for esterifying the 3-hydroxymethyl group of acephalosporin are known in the literature, e.g. see U.S. Pat. No.3,728,342, 3,532,694 and U.K. Pat. No. 1,365,954, and such methods maybe used with the novel 3-hydroxymethyl β-lactam derivatives of thepresent invention. The sulfonic acid esters may be formed by reaction ofthe 3-hydroxymethyl compound with a suitable sulfonic acid derivative,most preferably with a sulfonyl halide such as methanesulfonyl chloride,p-toluenesulfonyl chloride or trifluoromethanesulfonyl chloride in thepresence of an organic base. The acylating or esterifying agent used ispreferably one which will result in formation of the preferred3-esterified hydroxyl compounds mentioned above.

A compound of formula II' in which Z is etherified hydroxyl reducible byhydrogenolysis, e.g. benzyloxy, may be converted to the corresponding7-acylated 3-hydroxymethyl compound by catalytic hydrogenation accordingto methods known per se. Suitable hydrogenation catalysts include noblemetals, most preferably palladium or platinum and their oxides andhydroxides, and Raney nickel, said catalysts being optionally supportedon a conventional carrier such as carbon, diatomaceous earth, etc. Anespecially preferred catalyst is 20% Pd (OH)₂. Preferred solvents forthe hydrogenolysis reaction are non-reducible inert solvents such asmethanol, ethanol or ethyl acetate. The reaction is preferably conductedat atmospheric or slightly elevated pressure at room temperature. Whencarboxyl-protecting groups present in compound II' are reducible byhydrogenolysis, e.g. benzyl, p-nitrobenzyl, benzhydryl, etc., compoundII' may be simultaneously de-blocked and reduced to the desired3-hydroxymethyl free acid.

A compound of formula II' in which Z is hydroxyl may be converted to thecorresponding 7-acylated 3-etherified hydroxyl product by reacting thecompound with an etherifying agent by procedures used for cephalosporincompounds, e.g. those described in U.S. Pat. No. 3,665,003.

A compound of formula II' in which Z is hydroxyl may be converted to thecorresponding 3-heterocyclic ether compound by first forming a sulfonicacid ester at the 3-position, e.g. by reacting the 3-hydroxymethylcompound with a sulfonyl chloride in the presence of an organic base,and then displacing the sulfonate residue with a heterocyclic alcohol inthe presence of an organic base. The 3-heterocyclic ethers may also beprepared by nucleophilic displacement of the corresponding 3-halomethylcompound, the preparation of which is described below.

A compound of formula II' in which Z is acetoxy may be converted to thecorresponding 3-hydroxymethyl product by enzymatic hydrolysis, e.g. byuse of citrus acetyl esterase.

A compound of formula II' in which Z is hydroxyl may be converted to thecorresponding 3-halomethyl compound by reaction with a suitablehalogenating agent, e.g. a phosphorus halide such as phorphorustrichloride, phosphorus tribromide, phosphorus pentachloride, phosphoruspentabromide, phosphorus oxychloride, or phosphorus oxybromide. The3-iodomethyl compounds may also be formed by treating the 3-bromomethylor 3-chloromethyl compound with an alkali metal iodide.

Compounds of formula II' in which Z is hydroxyl may be converted to thecorresponding compounds having Z = OCONH₂ by reaction in an inertorganic solvent, e.g. benzene, with a source of cyanate ions, e.g., froman alkali metal cyanate, followed by treatment with trifluoroaceticacid. The cyanate ion and trifluoroacetic acid are preferably eachemployed in a molar ratio of about 2:1 with respect to the hydroxymethylstarting material.

A compound of formula II in the form of the free acid or a salt thereofmay be converted to a pharmaceutically acceptable salt thereof or to aphysiologically hydrolyzed ester or pharmaceutically acceptable saltthereof. Similarly, the product of formula II" in the form of an easilycleavable ester or salt thereof may be converted to the free acidproduct or a pharmaceutically acceptable salt thereof by removal of theesterifying group to form the free acid, e.g. by acidic or alkalinehydrolysis, by enzymatic hydrolysis (as with human or animal serum), byhydrogenolysis or by treatment with chemical reagents known to removeparticular blocking groups, e.g. sodium thiophenoxide as in U.S. Pat.No. 3,284,451, and subsequent treatment of the free acid with an acid orbase to form a pharmaceutically acceptable salt.

The easily cleavable esters of the compounds of formula II are useful asintermediates in the production of the free acid product. Thepivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethylesters are also useful as active antibacterial agents since an oraladministration they are rapidly hydrolyzed to the active metabolite.These esters are of particular interest because they provide an oraladministration different rates and amounts of absorption and givediffering concentrations of the active antibacterial agent in blood andtissues.

The 7-amino intermediates of general formula III may be prepared byselectively reducing a 7-azido intermediate of the formula ##STR54##wherein Z is halo, hydroxyl, etherified hydroxyl or hydroxyl esterifiedwith a carboxylic acid or sulfonic acid residue and R' is an easilycleavable ester carboxyl-protecting group. The carboxyl-protectedcompound may, if desired, be cleaved to produce the free-acidintermediate III which can be converted to a salt by methods known perse.

Preferred reducing agents for use in preparing the intermediates offormula III include chemical reducing agents such as zinc and ammoniumchloride, aluminum amalgam and hydrogen sulfide in the presence of abase, e.g. triethylamine or ammonia. Catalytic hydrogenation may also beemployed with such catalysts as noble metals, preferably platinum orpalladium including derivatives thereof such as oxides, hydroxides andhalides, or Raney nickel, said catalysts being optionally supported on aconventional carrier such as carbon or diatomaceous earth. Catalytichydrogenation is performed with a non-reducible inert solvent, e.g.methanol, ethanol or ethyl acetate, and preferably at atmospheric orslightly elevated pressure at room temperature.

Compound III in the carboxyl-protected form or a salt thereof may beused directly as a starting material in the N-acylation processdiscussed above. Alternatively, the protected intermediate may bede-blocked to form the free carboxylic acid which may then be optionallyconverted to a salt or to another carboxyl-protected form, e.g. aphysiologically hydrolyzed ester or salt thereof. By proper selection ofreduction conditions and protecting groups, azido intermediate IV' maybe converted either simultaneously or in stepwise fashion to the 7-aminofree acid III. Thus, if mild hydrogenation conditions are used, e.g.catalytic hydrogenation with 10% Pd-on-charcoal or a mild chemicalreducing agent such as H₂ S in the presence of a base such astriethylamine or ammonia, the azido group may be reduced withoutconcomitant removal of esters resistant to such conditions, e.g. benzylor p-nitrobenzyl. If stronger reducing conditions are used such as 30%Pd-on-diatomaceous earth, both the azido group and most reducible esterswill be simultaneously reduced. When it is desired to prepare anintermediate III where Z is etherified hydroxyl, it may be necessary toselect sufficiently mild reducing conditions, e.g. H₂ S with base, so asnot to affect a reducible ether moiety.

A preferred embodiment of the present invention is the processcomprising the consecutive steps of

1. selectively reducing a 7-azido intermediate of the formula IV' whereis halo, hydroxyl, hydroxyl esterified with a carboxylic acid orsulfonic acid residue or etherified hydroxyl and R' is an easilycleavable ester carboxyl-protecting group to produce acarboxyl-protected 7-amino intermediate of formula III and, if desired,removing the carboxyl-protecting group to produce the corresponding freeacid intermediate of formula III or optionally a salt thereof; and

2. N-acylating intermediate III or a salt thereof with an acylating acidof the formula R--COOH where R is an acyl group, or with its functionalequivalent as an acylating agent for a primary amine and, if desired,converting the so-produced product to the corresponding 7-acylatedproduct having a substituent Z as defined above different from that incompound III and, if desired, (a) where R" is a carboxyl-protectinggroup, converting the 7-acylated ester to the free acid compound or aphysiologically hydrolyzed ester or a pharmaceutically acceptable saltof said acid or ester, or (b) when R" is hydrogen, converting the7-acylated carboxylic acid to a physiologically hydrolyzed ester or apharmaceutically acceptable salt of said acid or ester and, if desired,resolving a resulting isomer mixture into its component isomers.

The 7β-azido intermediates IV' may be prepared by various methodsdepending on the nature of substituent Z. When Z is a group --OCOR₂derived from an acid R₂ COOH having a pK_(a) of between 3.5 to 5.5 andwherein R₂ is hydrogen, amino, (lower)alkyl, C₃ -C₇ cycloalkyl, C₃ -C₇cycloalkyl-(lower)alkyl, aryl aralkyl or aryloxyalkylene, said R₂ groupsbeing optionally substituted by one or more radicals selected from(lower)alkoxy, (lower)alkylthio, halogne, (lower)alkyl, nitro, hydroxy,acyloxy, carboxy, amino, (lower)alkylamino or acylanino, intermediatesof formula IV' may be prepared by cyclizing in an inert organic solventa compound of the formula ##STR55## wherein X and X' which may be thesame or different each represent a halogen atom, preferably bromine, oriodine and most preferably iodine, R' is an easily cleavable estercarboxyl-protecting group and Y' represents a suitable leaving group,preferably a group such as halo or sulfonyloxy, e.g. alkyl- orsubstituted alkylsulfonyloxy or aryl- or substituted arylsulfonyloxy andmost preferably a group selected from halo, --OSO₂ -(lower)alkylincluding especially --OSO₂ CH₃, --OSO₂ CF₃ and --OSO₂ C₆ H₄ CH₃, with abase selected from an anion of the formula R₂ COO⁻ derived from acarboxylic acid having a pK_(a) of between about 3.5 and 5.5 and inwhich R₂ is as defined above.

The dihalide starting material V may be used in either of its isomericforms ##STR56## or as a mixture of isomers. Formula V above is intendedto represent either of the individual isomers or the mixture. Anydihalide including a mixed dihalide, e.g. X = Cl, X' = Br, may be usedbut the most preferred compound is the diiodide. Compound V is reactedin an inert organic solvent, preferably a polar organic solvent such asdimethylformamide, with an excess of the base.

Any base derived from a carboxylic acid R₂ COOH satisfying the pK_(a)conditions above and wherein R₂ is as defined above may be employed inthe cyclization reaction. The preferred bases are the anions of theformula R₂ COO⁻ in which R₂ is hydrogen, (lower)alkyl and especiallymethyl, phenoxymethyl and tetrazolylmethyl. The most preferred bases arethe formate and acetate anions, e.g. from an alkali metal, ammonium orsubstituted ammonium formate or acetate.

The leaving group Y' should be one which is efficiently displaced underthe conditions of the base cyclization reaction. Suitable leaving groupsinclude halo and sulfonyloxy groups, i.e., alkyl- or substitutedalkylsulfonyloxy or aryl- or substituted arylsulfonyloxy. A mostpreferred Y' leaving group is the mesylate group.

Cyclization of compound V is conveniently carried out at roomtemperature or below.

Starting materials of formula V used in the above process may beprepared as described below in the section entitled "Preparation ofStarting Materials." Briefly summarized, the reaction scheme is as shownin Flow Sheet I: ##STR57##

Intermediates of formula IV' having Z either --OCHO or --OCOR₂ in whichR₂ is (lower)alkyl may be converted to the corresponding intermediatesof formula IV' wherein Z is hydroxyl by subjecting such compounds toacid hydrolysis, e.g. by treatment with a mineral acid in an aqueousacetone solvent system.

Intermediates of formula IV' in which Z is hydroxyl esterified with acarboxylic acid or sulfonic acid residue may be prepared by esterifyinga 3-hydroxymethyl intermediate of the formula ##STR58## with acarboxylic acid or sulfonic acid agent capable of introducing thedesired acyloxy or sulfonyloxy acyl moiety at the 3-position.Esterification in this embodiment of the present invention may beperformed in the same manner as described previously in connection withthe modification of the Z substituents in compounds of formula II. Apreferred embodiment comprises reacting an intermediate IV' in which Zis hydroxyl with acetic anhydride in the presence of an organic base toprepare the corresponding carboxyl-protected 3-acetoxymethyl 7-azidointermediate.

Another preferred embodiment involves esterifying intermediate IV' inwhich Z is hydroxyl with an esterifying derivative of methanesulfonicacid, trifluoromethanesulfonic acid or p-toluenesulfonic acid, e.g.methanesulfonyl chloride, p-toluenesulfonyl chloride or triflicanhydride, in the presence of an organic base, e.g. triethylamine, andin an inert organic solvent, e.g. methylene chloride, to produce anintermediate of formula IV' in which Z is methanesulfonyloxy,p-toluenesulfonyloxy or trifluoromethylsulfonyloxy.

Intermediates of formula IV' in which Z is hydroxyl may be converted tothe corresponding 3-halomethyl compounds by reaction with a halogenatingagent, e.g. a phosphorus halide such as phosphorus trichloride,phosphorus tribromide, phosphorus pentachloride, phosphoruspentabromide, phosphorus oxychloride or phosphorus oxybromide. The3-iodomethyl compounds may also be formed by treating the 3-bromomethylor 3-chloromethyl compound with an alkali metal iodide.

Intermediates of formula IV' in which Z is 3-halomethyl or 3-sulfonyloxymay be converted to the corresponding 3-heterocyclic ether intermediatesby nucleophilic displacement with a heterocyclic alcohol in the presenceof an organic base.

Carbamate intermediates of the formula ##STR59## may be prepared byreacting the corresponding carboxyl-protected 7-azido 3-hydroxymethylcompound in an inert organic solvent, e.g. benzene, with a source ofcyanate ions, e.g. an alkali metal cyanate, followed by treatment withtrifluoroacetic acid. The cyanate ion and trifluoroacetic acid arepreferably each employed in approximately a 2:1 molar ratio with respectto the azido starting material.

An alternate method provided by the present invention for preparation ofintermediates of formula IV' where Z is hydroxyl comprises cyclizingwith base in an inert organic solvent an intermediate of the formula##STR60## wherein R' is an easily cleavable ester carboxyl-protectinggroup and Y' represents a suitable leaving group, preferably a halo or asulfonyloxy group, e.g. alkyl- or substituted alkylsulfonyloxy or aryl-or substituted arylsulfonyloxy and most preferably a group selected fromhalo, --OSO₂ -(lower)alkyl including especially --OSO₂ CH₃, --OSO₂ CF₃and --OSO₂ C₆ H₄ CH₃.

The base used in the cyclization of VI may be selected from a widevariety of bases including especially bases of the following categories:

(a) anions derived from carboxylic acids having a pK_(a) of between 3.5and 5.5;

(b) tertiary organic amines such as a trialkylamine (e.g.,triethylamine), pyridine, N-methylpiperidine, N-methylmorpholine, etc.;

(c) alkali metal hydrides, e.g. sodium or potassium hydride; and

(d) organolithium compounds including especially lithium alkyls, e.g.,methyl lithium of butyl lithium.

Most preferred cyclization bases are the acetate and formate anions,e.g., from the alkali metal, ammonium or substituted ammonium formatesor acetates. The most preferred base is the acetate anion. The base ispreferably used in a molar excess relative to compound VI, and thereaction is conducted in an inert organic solvent, preferably a polarorganic solvent such as dimethylsulfoxide or dimethylformamide. Thetemperature for the cyclization is not critical, and room temperaturemay be used for convenience.

Compound VI used in the above process may be prepared by the proceduresdescribed below in the section entitled "Preparation of StartingMaterials". A summary of the reaction scheme is shown in Flow Sheet 2:##STR61##

The 7β-azido intermediates of formula IV' where Z is etherified hydroxylmay be prepared by cyclizing with base in an inert organic solvent anenol intermediate of the formula ##STR62## wherein Z is an etherifiedhydroxyl group, R' is an easily cleavable ester carboxyl-protectinggroup and Y' represents a suitable leaving group, preferably a halo orsulfonyloxy, e.g. alkyl- or substituted alkylsulfonyloxy or aryl- orsubstituted arylsulfonyloxy and most preferably a group selected fromhalo, --OSO₂ -(lower)alkyl including especially --OSO₂ CH₃, --OSO₂ CF₃and --OSO₂ C₆ H₅ CH₃.

The base used to cyclize enol VII may be any of the bases mentionedabove in connection with the cyclization of intermediate VI and ispreferably an acetate or formate anion and most preferably the acetateanion, e.g., from an alkali metal, ammonium or substituted ammoniumacetate. Cyclization is carried out in an inert organic solvent, e.g.,methylene chloride, dimethylformamide or dimethylsulfoxide, with anexcess of base, and may conveniently be done either at room temperatureor under reflux.

Compound VI used in the above process may be prepared by the generalmethod described below in the section entitled "Preparation of StartingMaterials". A summary of the reaction scheme is shown in Flow Sheet 3:##STR63##

The pharmaceutically active compounds of the present invention arepotent antibacterial agents useful in the treatment of infectiousdiseases in poultry and animals, including man, caused by manyGram-positive and Gram-negative bacteria. The active compounds are alsoof value as nutritional supplements in animal feeds and as agents forthe treatment of mastitis in cattle.

The novel medicaments provided by the present invention may beformulated as pharmaceutical compositions comprising, in addition to theactive ingredient, a pharmaceutically acceptable carrier or diluent. Thecompounds may be administered both orally and parenterally. Thepharmaceutical preparations may be in solid form such as capsules,tablets or dragees, or in liquid form such as solutions, suspensions oremulsions. In the treatment of bacterial infections in man, the activecompounds of this invention may be administered parenterally or orallyin an amount of from about 5 to 200 mg/Kg./day and preferably about 5 to20 mg./Kg./day in divided dosage, e.g. three or four times a day. Theyare administered in dosage units containing, for example, 125, 250 or500 mg. of active ingredient with suitable physiologically acceptablecarriers or excipients.

Illustrative examples of the preparation of starting materials andcompounds of the present invention follow. These examples are given inillustration of, but not in limitation of, the present invention. Alltemperatures are in degrees Centigrade. DMF representsdimethylformamide, THF stands for tetrahydrofuran and EEDQ is the amidebond forming reagent having the structure ##STR64##

The 7-acylamido compounds prepared in the examples which follow all havethe hydrogen atoms at carbons 6 and 7 cis with respect to each otherand, unless indicated, the products are racemic mixtures in the sensethat they are composed of equal parts of the two isomers having thefollowing structures: ##STR65##

Preparation Of Starting Materials

1. Preparation of intermediates of formulae ##STR66##

Benzyl Oximino-Acetoacetate ##STR67##

The procedure was essentially the same as that used to make thecorresponding ethyl ester by H. Adkins and J. Reeve, JACS 60, 1328(1938).

In a three necked one liter flask fitted with a thermometer, a droppingfunnel and a magnetic stirrer were placed 173 g. (0.9 mole) of benzylacetoacetate [The benzyl acetoacetate was prepared as described by Bakeret al., J. Org. Chem. 17, 91 (1952)] and 130 ml. of glacial acetic acid.The contents were cooled in an ice bath and a solution of 69 g. (1 mole)of sodium nitrite in 130 ml. of water was added over a period of half anhour; the temperature was kept at 0° to 10° C. After the reactionmixture was stirred for 1 hour at room temperature, 400 ml. of water wasadded and the stirring was continued for an additional two hours. Thereaction mixture was extracted three times with 200 ml. portions ofdiethyl ether. The diethyl ether extracts were combined, washed oncewith water, three times with saturated sodium bicarbonate solution andonce with brine. After drying over anhydrous sodium sulfate, the diethylether solution was evaporated leaving [1.1] as a clear oil whichsolidified upon trituration with petroleum ether (30°-60°) to give 186.5g. (93.2%) of white solid. Its NMR spectrum was consistent with theassigned structure. Generally the product was used as such in subsequentreaction but it can be recrystallized from toluene, m.p. 81°-82° C.

Benzyl Oximino-Acetoacetate Ethylene Ketal ##STR68##

In a 2 liter flask fitted with a Dean Stark water separator and acondenser were placed 186.5 g. (0.85 mole) of benzyloximino-acetoacetate [1.1], 62 g. (1mole) of ethylene glycol, 800 ml. ofbenzene (reagent grade) and 2 g. (10.5 mmole) of p-toluenesulfonic acidmonohydrate. The reaction mixture was boiled at reflux until 15 ml. ofwater was removed (3 hours). The benzene solution was washed once withsaturated sodium bicarbonate solution and once with brine. After dryingover anhydrous sodium sulfate, the benzene solution was evaporated,leaving 212 g. (94% yield) of benzyl oximinoacetoacetate ethylene ketal[2.1] as a light yellow oil! Its NMR spectrum was consistent with theassigned structure. Generally, the product was used as such insubsequent reactions but one of the isomers can be crystallized² fromtoluene-petroleum either (b.p. 30°-60° C.); m.p. 52° C.

Anal. Calc'd. for C₁₃ H₁₅ NO₅ : C, 58.86; H, 5.70; N, 5.28. Found: C,58.97; H, 5.68; N, 5.12.

1. A mixture of the syn and anti isomers.

2. Only 35% of the oil could be crystallized.

Benzyl Amino-Acetoacetate Ethylene Ketal ##STR69##

Freshly prepared aluminum amalgam¹ (from 27 g. of aluminum foil) in athree-necked one liter flask was covered with 500 ml. of diethyl ether.The flask was fitted with a mechanical stirrer, a condenser, and adropping funnel. A solution of benzyl oximino-acetoacetate ethyleneketal [2.1] (132.5 g.; 0.5 mole) in 300 ml. of wet diethylether² wasadded dropwise at such a rate as to maintain boiling at reflux. Afterstirring for four hours, the reaction mixture was filtered through aBuchner funnel. The filtrate was evaporated leaving 110 g. of yellowishoil. The oil was dissolved in 800 ml. of dry diethylether and dryhydrogen chloride gas was bubbled into the solution until no furtherprecipitation occurred. The white precipitate was filtered off andwashed once with diethylether and then dried in vacuo. This provided 108g. of benzyl aminoacetoacetate ethylene ketal hydrochloride³ [3.1]; m.p.157°-158° C.

Anal.Calc'd. for C₁₃ H₁₇ NO₄.HC1: C, 54.26; H, 6.31; N, 4.87. Found: C,53.96; H, 6.19; N, 4.60.

To obtain the free base, the hydrochloride salt was suspended in 500 ml.of diethylether and concentrated ammonium hydroxide was added withshaking until the solid went into solution. The diethylether layer wasseparated and washed twice with brine. After drying over anhydroussodium sulfate, the solvent was evaporated leaving 90 g. (71% yield) ofcolorless oil.

Schiff Base Formation and β-Lactam Formation ##STR70##

In a one liter flask fitted with a Dean Stark water separator and acondenser were placed 70.3 g. (0.28 mole) benzyl aminoacetoacetateethylene ketal [3.1], 37 g. 80.28 mole) cinnamaldehyde, and 750 ml. ofmethylene chloride (reagent grade). The mixture was boiled at reflux for30 minutes and then 400 ml. of methylene chloride were distilled off.The concentrated solution was then dried over anhydrous sodium sulfateand then evaporated to dryness in vacuo¹. The residual oil was checkedby NMR to ensure that Schiff base formation was complete beforecontinuing on to the next step.

The freshly prepared Schiff base [4.1] was diluted with 600 ml. ofmethylene chloride² and cooled to 0° C. (ice-salt bath). Triethylamine(31.1 g.; 0.308 mole) was added and then a solution of 36.2 g. (0.308mole) of azidoacetyl chloride³ in 362 ml. of methylene chloride² wasadded dropwise at 0° C. over a period of one hour. The reaction mixturewas stirred for an additional hour at room temperature⁴ and thenevaporated on a rotary evaporator at reduced pressure while being heatedon a 35° C. water bath⁵. The residue was diluted with 500 ml. ofdiethylether and filtered. The filtrate was washed twice with brine anddried over anhydrous sodium sulfate. Evaporation of this solutionyielded 117.5 g. (94% yield) of styryl β-lactam [5.1]. Its NMR and IRspectra are consistent with the assigned structure and indicate thepresence of a mixture of isomers, diasteriomeric at the carbon α to thecarbonyl of the benzyl ester.

Styryl β-lactam [5.1] (117.5 g.; 0.262 mole) was dissolved in one literof methylene chloride (reagent grade), cooled to -50° to -60° C. in adry ice-acetone bath, and ozonized until a faint blue-green colorappeared. The solution was then flushed with nitrogen until the colorfaded. Methylsulfide (100 ml.) was added to the -50° C. solution, whichwas then allowed to slowly reach 25° as the cooling bath graduallymelted. It was kept overnite at room temperature under nitrogen and thenit was washed twice with 1% sodium bicarbonate solution, twice withbrine, dried over anhydrous sodium sulfate, and evaporated to dryness.The resulting oil triturated four times with 100 ml. portions ofpetroleum ether (b.p. 30°-60° C.) to remove benzaldehyde. The oil wasthen triturated carefully with diethylether whereupon it solidified. Thesolid was filtered off and dried to provide 75 g. (71.5%) of aldehyde[6.1] as a mixture of isomers diasteriomeric at the carbon α to thecarbonyl of the benzyl ester. Recrystallization of [6.1] from ether gavewhite crystals, m.p. 101°-102° C. (corrected).

Anal. Calc'd. for C₁₇ H₁₈ N₄ O₆ : C, 54.54; H, 4.84; N, 14.96. Found: C,54.75; H, 4.87; N, 14.89. ##STR72##

The aldehyde [6.1] (116.3 g.; 0.31 mole) was dissolved in 600 ml. of THF(reagent grade) and the solution was then cooled to -10° C.(ice-methanol bath). Sodium borohydride (5.88 g.; 0.155 mole) was addedand the reaction mixture was stirred 1 hour. 10% aqueous hydrochloricacid was added until the mixture was slightly acidic, then 600 ml. brinewas added. The THF layer was separated and the aqueous phase wasextracted twice with 250 ml. portions of diethylether. The combinedorganic phases were washed twice with 400 ml. portions of brine, driedover anhydrous sodium sulfate, and evapoated in vacuo to yield 117.3 g.of crude alcohol [7.1] as an orange oil. This oil was used as such inthe next reaction.

A solution of methanesulfonyl chloride (37.8 g.; 0.34 mole) in 100 ml.of methylene chloride¹ was added dropwise at 0° C. (ice-water bath) to astirring solution of alcohol [7.1] (105.6 g.; 0.28 mole, triethylamine(56.6 g.; 0.34 mole) and one liter of methylene chloride¹. Afterwards,the reaction was stirred for 30 hours at 25° C. It was then washed twicewith brine (500 ml. portions), dried over anhydrous sodium sulfate, andevaporated in vacuo. The resulting oil was dissolved in methylenechloride, treated with norite, and then filtered over approximately 200g. of activity I silica gel. The silica gel was then washed withapproximately 2 liters of methylene chloride. The filtrate wasevaporated to dryness and the resulting oil (116 g.) was covered withdiethylether. It crystallized on standing to give 87.2 g. (80% from[6.1] of mesylate [8.1] as offwhite solid, m.p. 97°-99° C (corrected).

A mixture of mesylate [8.1] (3.19 g.; 6.43 mmole) and 30 ml. of 95%trifluoroacetic acid was stirred at 25° for 2 hours. The mixture wasdiluted with 300 ml. of brine and extracted three times with methylenechloride (100 ml. portions). The combined extracts were washed threetimes with water (50 ml. portions, until neutral), dried (anhydroussodium sulfate) and evaporated to dryness in vacuo leaving 3.17 g. of abrown oil. NMR spectra of this oil indicate the presence of >90% enol[9.1].

Crude enol [9.1] (48.0 g.; 0.117 mole) and triflic anhydride (33.0 g.;0.117 mole) were dissolved in 500 ml. of methylene chloride and thesolution was then cooled to 0° C. (ice-water bath). A solution oftriethylamine (11.8 g.; 0.117 mole) in 80 ml. of methylene chloride² wasadded dropwise over a period of 40 minutes. When the addition wascomplete, the ice-water bath was removed and the mixture was stirred at25° for 45 minutes. The mixture was then poured into 300 ml. of icewater and washed with cold water until the pH of the washings wasapproximately 6. The extract was dried (anhydrous sodium sulfate) andevaporated in vacuo to give 54.0 g. of crude triflate [10.1] as a darkred oil. This oil was dissolved in 400 ml. of benzene (USP) and passedthrough a 11/2 pad of activity III silica gel. The pad was washed with 1l of benzene. Evaporation of the benzene gave 38.3 g. of a yellow oil.This oil was carefuly triturated with 50 ml. of absolute ethanol andthen cooled at 0° C. for 2 hours. The resulting white solid was filteredoff and dried in vacuo to give 19.5 g. of triflate [10.1] as one isomer,m.p. 57°-59° C. (corrected).

Anal. Calc'd. for C₁₇ H₁₇ F₃ N₄ O₄ S₂ : C, 37.67; H, 3.14; N, 10.33; S,11.82. Found: C, 37.40; H, 3.12. N, 10.43; S, 11.73.

Triethylamine (1 g.; 0.01 mole) was added to a stirred solution oftriflate [10.1] (5.42 g.; 0.01 mole) in 55 ml. of methylene chloride(A.R.) at room temperature. After stirring for five minutes (at whichpoint TLC shows complete formation of allene [11.1], a solution ofbromine (10 ml. of 1M solution in CCl₄ ; 0.01 mole) was added dropwise.After addition of the bromine, the mixture was concentrated, absorbedonto Activity I silica gel and dry column chromatographed on Activity Isilica gel by eluting with methylene chloride (USP). This yielded onefraction (uniformly one spot by TLC) weighing 2.5 g. (45%). Its IR, UV,and NMR spectra were consistent with the expected dibromide structure[13.1].

Anal. Calc'd. for C₁₆ H₁₂ Br₂ N₄ O₆ S: C, 34.80; H, 2.92; N, 10.15.Found: C, 35.25; H, 2.97; N, 10.02.

1A. Preparation of intermediates of formulae ##STR75##

A solution of triethylamine (101 mg., 1.00 mmole) in 1.4 ml. ofmethylene chloride was added with stirring to a solution of triflate[10.1] (542 mg., 1.00 mmole) in 5.4 ml. of methylene chloride at 0° C.After allowing the solution to warm to 24° over 15 minutes, a solutionof iodine (254 mg., 1.00 mmole) in 7.5 ml. of methylene chloride wasadded with stirring over 30 minutes, then washed with water, dried,decolorized, filtered and the solvent evaporated in vacuo to give thediiodide [23.1] (588 mg.; 91% yield) in greater than 95% purity. The IRand NMR spectra were consistent for the proposed structures.

Anal. Calc'd. for C₁₆ H₁₆ N₄ O₆ I₂ S: C, 29.74; H, 2.50; N, 8.67; I,39.28; S, 4.96. Found: C, 29.76; H, 2.47; N, 8.61; I, 39.37; S, 5.18.

2. Preparation of intermediates of formula ##STR76##

Butyl lithium (1.1 moles, 2.4 M in hexane) was added under an atmosphereof dry nitrogen to 1 liter of dry tetrahydrofuran (freshly distilledfrom lithium aluminum hydride) at -60° C. It was followed by thedropwise addition of isopropylcyclohexylamine (dried over potassiumhydroxide pellets) (200 ml., 1.1 moles) and then benzyl acetate (165 g.,1.1 moles). After stirring for 3/4 hour at -60° C., it was treatedfairly rapidly with diethoxy ethyl acetate [1.2]¹ (178 g., 1.01 moles).The mixture was allowed to come slowly to 20° C. It was then cooled to0° C. and acidified with 10% hydrochloric acid. Ether was added and theether extract was washed with water (twice) and then brine. It was driedover anhydrous sodium sulfate and evaporated to give 283 g. (100%) ofβ-keto ester [2.2] of suitable purity to be used directly.

A solution of the β-keto ester [2.2] (283 g., 1.01 moles) in glacialacetic acid (300 ml.) was cooled to 0° C. and treated dropwise below 5°C. with a solution of sodium nitrite (100 g., 1.45 moles) in 250 ml. ofwater. By the end of the addition, the solid product has begun to form.After coming to room temperature over 11/2 hours, it was treated slowlywith 400 ml. of water. The solid was collected by filtration and washedwith cold water.

This solid was then dissolved in 1 liter of ether and washed with waterand brine. It was dried over anhydrous sodium sulfate and evaporated togive 288 g. (93%) of the oxime [3.2] which was used without furtherpurification.

For analysis, a sample was crystallized twice from ether/petroleum ether(b.p. 30°-60°) giving white needles with m.p. 95°-97° C.

Anal. Calc'd. for C₁₅ H₁₉ NO₆ : C, 58.24; H, 6.19; N, 4.53. Found: C,58.28; H, 6.37; N, 4.51. ##STR78##

A solution of the keto-oxime [3.2] (288 g., 0.94 moles) in 1 liter oftetrahydrofuran was cooled to 0° C. and treated portionwise with sodiumborohydride (17.8 g., 0.47 moles). After stirring for 3/4 hours at 0°C., it was allowed to come to room temperature over 11/2 hours.

It was then recooled to 0° C., acidified carefully with dilutehydrochloric acid, and extracted with ether. The ether extract waswashed with water and brine. It was dried over anhydrous sodium sulfateand evaporated to give 241.5 g. (83%) of the alcohol [4.2] which wasused without purification. ##STR79##

A solution of the alcohol [4.2] (241.5 g., 0.78 moles) in 1 liter ofether was treated with dihydropyran (130 g.; 1.5 moles) and then with4.0 g. of p-toluenesulfonic acid monohydrate. Some cooling was requiredto keep the initial reaction at gentle reflux. When the initial reactionhad ceased, it was stirred at room temperature for 2 hours.

It was then poured onto 500 ml. of saturated sodium bicarbonate solutionand the ether extract was washed with water and brine. It was dried overanhydrous sodium sulfate and evaporated to give 356 g. of the THPprotected alcohol [5.2]¹. It was used as such in the following reaction.

A solution of the oxime [5.2] (64 g.; 0.143 moles) in tetrahydrofuran(200 ml.) was added to a slurry of aluminum amalgam (prepared from 0.72moles of aluminum foil according to the method of Vogel, "PracticalOrganic Chemistry", p. 193-98) at such a rate as to maintain a vigorousreaction. After stirring for 3-4 hours, a fresh amount of aluminumamalagam (0.36 moles) was added at such a rate as to maintain a vigorousreaction. After stirring for 16 hours, it was filtered through celitewashing well with ether.

The filtrate was evaporated and the residue was dissolved in ether andtreated with a solution of oxalic acid (12.9 g., 0.143 moles) in ether.The mixture was then extracted with cold water twice and the combinedaqueous extracts were acidified with concentrated ammonium hydroxide inthe cold. It was then extracted with ether and the extract dried overanhydrous sodium sulfate. Evaporation gave 22.8 g. (42%) of the amine[7.2]. ##STR81##

The amine [7.2] (28.9 g.; 0.076 moles) was treated withtrans-cinnamaldehyde (10.0 g.; 0.076 moles) in methylene chloride (200ml.) and refluxed for 2 hours. It was then dried over anhydrous sodiumsulfate for several minutes and evaporated. The residue was dissolved ina further 200 ml. of methylene chloride and then evaporated again. Thisprocedure was repeated twice more giving a residue with NMR showingalmost complete Schiff base [8.2] formation.

The Schiff base [8.2] was immediately dissolved in methylene chloride(200 ml.) (dried over molecular sieves) and cooled to 0° C. It wastreated with triethylamine (dried over potassium hydroxide pellets) (7.7g.; 0.076 moles) and then dropwise at 0° to 5° C. with a solution ofazidoacetyl chloride (9.1 g.; 0.076 moles) in dry methylene chloride(100 ml.) under an atmosphere of dry nitrogen. When the addition wascomplete, it was allowed to come to room temperature for 16 hours andthen refluxed for 3/4 hour.

On cooling, the solution was washed with water, 1% hydrochloric acid,and brine. It was dried over anhydrous sodium sulfate and evaporated togive 43.0 g. (100%) of crude styryl compound [9.2]. ##STR82##

The crude styryl compound [9.2] (14.4 g.; 0.025 moles) was dissolved inmethylene chloride (80 ml.) and cooled to -60° C. in a dry ice:isopropanol bath. Ozone was then bubbled through the solution until theappearance of a faint blue color persisted. It was then flushed withoxygen to remove excess ozone and treated with methyl sulfide (10 ml.)to decompose the ozonide. It was allowed to come to room temperatureover 4 hours and then washed with water, 5% sodium bicarbonate, water,and brine. It was dried over anhydrous sodium sulfate and evaporated.The residue was evaporated under high vacuum at 40°-50° C. for 18 hoursto remove most of the benzaldehyde. The residue (12.1 g.) had NMRshowing about 50-60% of free aldehyde [10.2] by integration. It was usedas such in the next step. ##STR83##

The crude aldehyde [10.2] (12.1 g.; 0.024 moles) was dissolved intetrahydrofuran (100 ml.) and cooled to 0° C. It was treated portionwisewith powdered sodium borohydride (0.46 g.; 0.012 moles) over a shortperiod of time. After 1/2 hour stirring at 0° C., it was poured ontoice-cold 10% acetic acid and the acidified mixture was extracted withether.

The ether extract was washed with water (twice), 10% sodium bicarbonate,and brine. It was dried over anhydrous sodium sulfate and evaporated togive 10.9 g. of crude alcohol [11.2]. ##STR84##

The crude alcohol [11.2] (12.8 g.) was dissolved in methylene chloride(100 ml.) (dried over molecular sieves) and cooled to 0° C. It wastreated with dry triethylamine (2.23 g.; 0.022 moles) and then dropwisewith a solution of methanesulfonyl chloride (2.51 g.; 0.022 moles) indry methylene chloride (30 ml.).

It was allowed to come to room temperature over 3 hours and then washedwith water, 1% hydrochloric acid, and brine. It was dried over anhydroussodium sulfate and evaporated to give 11.8 g. of crude mesylated alcohol[12.2]. ##STR85##

The crude THP-protected alcohol [12.2] (11.8 g.) was dissolved intetrahydrofuran (100 ml.) and treated with cooling (keeping thetemperature below 15° C.) with 18% hydrochloric acid (50 ml.). Afterstirring for an additional hour in the cold, thin layer chromatographyshowed loss of the starting material.

It was diluted with water and extracted with ether. The ether extractwas washed with water and brine. It was dried over anhydrous sodiumsulfate and evaporated.

The crude residue was purified by dry-column chromatography on silicagel (Activity III) using ether as the solvent. This gave 3.5 g. of theacetal [13.2]. The yield is 28% overall from the amine [7.2]. ##STR86##

The acetal [13.2] (1.5 g.; 3 mmoles) was stirred for 45 minutes with 5ml. of 95% trifluoroacetic acid and then treated with methylene chlorideand water. The methylene chloride extract was washed with water, 10%sodium bicarbonate, and brine. It was dried over anhydrous sodiumsulfate and evaporated to give 1.0 g. (78%) of crude hydroxy aldehyde[14.2].

3. Preparation of intermediates of formula ##STR87##

Benzyl alcohol (6500 ml.) is added to a 12 liter 3-necked flask equippedwith an efficient stirrer, condenser and a heating mantle. The alcoholis heated above the melting point of sodium (˜100°-110° C.) and sodiummetal (600 g., 26 moles) is added in small pieces with vigorous stirringat a rate sufficient to maintain a gentle reflux and to prevent theaccumulation of large amounts of sodium in the reaction flask (˜3-4hours). The solution is then cooled to about 80° C. and chloroaceticacid [1.3] (1120 g.; 12 moles) dissolved in the minimum amount of benzylalcohol is added dropwise fairly rapidly. The mixture is heated underreflux for 4 hours, cooled to room temperature and diluted with 10liters of ether. The precipitated sodium salt of benzyloxyacetic acid iscollected by filtration, washed with ether, dissolved in cold water andacidified to pH 3 with concentrated HCl. Extraction with CH₂ Cl₂, dryingand concentration gives 1952 g. (98% yield) of benzyloxyacetic acid[2.3] as a crude oil which is esterified without further purification.If desired, the acid can be purified by distillation: b.p. 130°-132°C./0.1 mm. ##STR88##

Ethanol (7 liters) in a 12 liter 1-neck flask equipped with condenserand drying tube is saturated with dry HCl. Benzyloxyacetic acid [2.3](1952 g., 11.8 mole) is added in one portion and the solution is heatedunder reflux for 5 hours. Most of the ethanol is removed by distillationat normal pressure. The residue is cooled to about 5° C. and dilutedwith about 4 l. of cold water. Extraction with ether, washing of theether with cold water, saturated NaHCO₃ solution and again with coldwater, drying over Na₂ SO₄ and concentration yields an oil which ispurified by distillation to give 1425 g. (62% yield) of [3.3]; b.p.138°-140° C./8 mm. The NMR spectrum indicated a benzyl --CH₂ : singletat 4.57 δ, aromatic H at 7.3 δ, O--CH₂ --CO- at 4.07 δ and an ethylquartet and triplet centered at 4.18 and 1.25 δ. ##STR89##

All operations are carried out under an atmosphere of dried N₂. Areaction flask (12 l. 3-neck flask with stirrer, N₂ inlet, 500 ml.addition funnel and cold temperature thermometer) is charged with 3 l.of dried tetrahydrofuran and cooled to -78° C. in a bath of dryice-acetone. Three bottles of Buli (2.4M in hexane, 1 mole/bottle) areadded via a flexible adaptor under N₂. N,N- isopropyl cyclohexylamine(423.8 g., 3 mole) is added via the addition funnel over a period ofabout 15 minutes and stirring is continued for 15 minutes. Benzylacetate (450.5 g., 3 mole) is then added dropwise over a 30 minuteperiod and the solution is stirred for 30 minutes. Ethylbenzyloxyacetate [3.3] (555.0 g., 2.85 mole) is added dropwise overabout 30 minutes and the cooling bath is removed at the end of theaddition. The reaction mixture is stirred and allowed to come to roomtemperature in about 2 hours. When the internal temperature reaches0°-5°, a voluminous solid separates. Anhydrous ether (about 3 l.) isthen added to complete the precipitation. The solid is collected byfiltration and washed with ether on the filter.

The solid is added to a vigorously stirred mixture of 250 ml. ofconcentrated HCl, 500 ml. of water, 500 ml. of ice and 3 l. of ether.When solution is complete the phases are decanted and the aqueous phaseis extracted once more with ether. The combined ether layers are washedwith brine, dried over Na₂ SO₄ and concentrated to leave 659 g. (78%) ofoily benzyl γ-benzyloxyacetoacetate. [4.3]: TLC: (silica gel, ether-pet.ether 2:1): 1 spot, Rf. 0.5, NMR indicates (aromatic H at 7.23δ; benzylester CH₂, singlet at 5.05δ; benzyl ether CH₂, singlet at 4.42δ; O--CH₂CO, singlet at 4.0δ; and COCH₂ CO₂, singlet at 3.48δ.

This oil is used without purification in the next step. Distillation oflarge amounts proceeds with extensive decomposition but a small amount(5 ml.) can be purified by distillation if necessary: b.p.180°-182°/0.05. The TLC and NMR of the crude product and of a distilledsample are identical. New spots appear on TLC if this oil is stored forsome time. ##STR90##

A solution of sodium nitrite (165 g., 2.4 mole) in 660 ml. of water isadded to the cooled (˜10° C.) solution of benzyl γ-benzyloxyacetate fromthe step immediately above in 1100 ml. of acetic acid. The rate ofaddition is regulated so that the temperature of the reaction mixturedoes not exceed 25° C. A white solid crystallizes out when aboutone-half of the nitrite solution is added. After the end of the additionthe mixture is stirred for an additional 30 minutes at 10° and dilutedslowly with 2 l. of water. The solid oxime is collected by filtrationwashed with cold water until the washings are neutral, and dried invacuo (16 hours) to give 856 g. of oxime as a white solid. The oxime isdissolved in methylene chloride, the water removed by decantation, theorganic phase washed with brine, dried over Na₂ SO₄ and concentrateduntil crystals start appearing. Crystallization is completed by dilutionwith petroleum ether (30°-60°). Filtration, washing with petroleum etherand drying affords 585 g. (81%) of oxime [5.3]; m.p. 92°-95°, NMRindicates aromatic H at 7.27 and 7.30 δ; benzyl ester CH₂ at 5.27δ andether CH₂ at 4.5δ. This material is used in the next step.

An analytical sample prepared in benzene-petroleum ether had thefollowing properties: m.p. 96°-97°.

Calc'd for C₁₇ H₁₈ NO₅ : C, 66.05; H, 5.23; N, 4.28. Found: C, 66.06; H,5.25; N, 4.23. ##STR91##

The keto-oxime [5.3] (400 g., 1.22 mole) dissolved in 1000 ml. dioxaneis cooled to 5°, stirred vigorously and treted portionwise with finelypowdered sodium borohydride (23.4 g., 0.62 mole) over a 30 minuteperiod. The temperature remains at 10° for a while and then a vigorousreaction sets in which must be controlled with an ice-salt bath(temperature goes up to 50°-60°). After 1 hour TLC (silica gel,ether-petroleum ether) shows disappearance of the starting material. Themixture is stirred an additional hour, poured into ice-cold dilutehydrochloric acid and extracted with ether (2 × 1 l.). The combinedextracts are washed with water (5 × 1 l.) and with brine. Drying andconcentration leaves a tan-colored solid which is recrystallized inbenzene-petroleum ether to give 308 g. (77% yield) of the alcohol-oxime[6.3] as a white solid, m.p. 83°-85°. NMR indicates aromatic H at 7.23δ, ester CH₂ at 5.18δ, benzyl ether at 4.48δ, (triplet); H of CHOH at4.67δ, Ch₂ ether at 3.65δ and exchanageable H at 3.42 and 10.05δ.

Calc'd for C₁₂ H₁₉ NO₅ : C, 65.64; H, 5.81; N, 4.25. Found: C, 65.62; H,5.91; N, 4.26. ##STR92##

To a well-stirred suspension of hydroxy-oxime [6.3] (298 g., 0.91 mole)in dihydropyran (650 ml., 7.1 mole) are added 15 to 20 drops ofconcentrated HCl and the mixture is stirred at room temperature. After1/2 hour a clear solution is obtained and after 4 hours TLC indicatesdisappearance of the starting hydroxy-oxime. The solution is dilutedwith technical ether (1.5 l.) and poured into a 10% NaHCO₃ solution. Theether phase is washed with brine, dried and concentrated to leave 402 g.of a yellow oil which consists of a mixture of mono andbis-tetrahydropyranyl derivatives.

The above oil (394 g.) dissolved in ether (technical 600 ml.) is addeddropwise to freshly prepared aluminum amalgam covered with technicalether (about 1000 ml.) (the amalgam is prepared using 120 g. aluminumfoil as described in Vogel, Practical Organic Chemistry) at a ratesufficient to maintain a vigorous reflux. The mixture is stirred 2 hoursafter the end of the addition. After removal of the insoluble materialsby filtration over Celite, the filtrates are dried over Na₂ SO₄ andtreated with anhydrous oxalic acid (72 g., 0.8 mole) dissolved in theminimum volume of ethanol. After two hours at 0°-5° the solid oxalatesalt is collected by filtration, washed with ether and dried to give 215g. of white solid, m.p. 115°-119°. This crude oxalate is used forregeneration of the free base.

An analytical sample of the oxalate, prepared in methanol-ether gives:m.p. 136°-137°.

Calc'd for C₂₃ H₂₉ NO₅.C₂ H₂ O₄ : C, 61.34; H, 6.38; N, 2.86. Found: C,61.25; H, 6.50; N, 2.76.

The free base [8.3] is regenerated by adding the solid oxalate to a wellstirred mixture of an excess of ice-cold concentrated ammonium hydroxideand ether: 152 g. (42%), yellowish heavy oil. NMR indicates aromatics at7.30δ, benzyl ester CH₂ at 5.1δ, benzyl ether CH₂ at 4.47δ and NH₂ at1.82δ. ##STR93##

A mixture of the amine [8.3] (117.2 g.; 0.294 mole) and cinnamaldehyde(38.8 g.; 0.294 mole) in 500 ml. of reagent grade CH₂ Cl₂ was refluxedfor one hour. The solvent was removed on the rotary evaporator and theresidue redissolved in 500 ml. of fresh CH₂ Cl₂. Part of the solvent wasdistilled at atmospheric pressure (350 ml.) and the remainder was takento dryness on the rotary evaporator to give an oil consisting of crude[9.3].

The above oil was dissolved in 400 ml. of CH₂ Cl₂, triethylamine (45ml., 0.32 mole) was added and the solution was cooled to 0° C. Asolution of azidoacetyl chloride (38.4 g.; 0.32 mole) in 200 ml. ofmethylene chloride was added to the above cooled and stirred solution in1.5 hour. The mixture was allowed to come slowly to room temperature andwas stirred for 16 hours. It was then heated under reflux for 1 hour,cooled, washed with ice-cold water, with ice-cold saturated sodiumbicarbonate solution and with ice-cold brine solution, dried over Na₂SO₄ and concentrated on the rotary evaporator to leave 171.2 g (98%) ofan oil whose IR and NMR spectra are consistent with structure [10.3].

If needed, purification can be effected through chromatography over acolumn of silica gel. The crude product is normally used in the nextstep. ##STR94##

A solution of the crude styryl compound [10.3] (117.6 g., 0.198 mole) in600 ml. of CH₂ Cl₂ was cooled to -60° C. and ozonized until a faint bluecolor appeared. Dimethyl sulfide (75 ml.) was added and the solution wasallowed to come slowly to room temperature. After 18 hours, the solutionwas washed several times with ice-cold water, dried over Na₂ SO₄ andconcentrated on the rotary evaporator to leave 104 g. of an oil. Thepure aldehyde [11.3] (34.6 g.) was obtained in 32% yield afterchromatography on 1200 g. of activity 2 silica gel using the dry columntechnique and eluting with ether-petroleum ether. ##STR95##

The pure aldehyde [11.3] (35.5 g., 0.068 mole) was dissolved in 250 ml.of reagent grade THF, cooled to -5° C., and treated portionwise undergood stirring with sodium borohydride (1.29 g., 0.034 mole). Afterone-half hour, the mixture was acidified with ice-cold 10% acetic acidand was extracted with ether. The combined ether extracts were washedwith ice-cold water and ice-cold 1% NaHCO₃ solution, dried andconcentrated to leave 33.4 g. (94%) of crude alcohol as an oil whose NMRspectrum is consistent with structure [12.3].

The alcohol was purified by column chromatography over silica gel.##STR96##

The purified alcohol [12.3] (15 g., 0.0286 mole) was dissolved in 100ml. of CH₂ Cl₂, cooled to 0° ., treated with triethylamine (4.2 ml.,0.03 mole), and treated dropwise with methanesulfonyl chloride (3.44 g.,0.03 mole) dissolved in 50 ml. of CH₂ Cl₂. After 2 hours, the mixturewas washed with ice-cold water and ice-cold brine solution, dried andconcentrated on the rotary evaporator to give 15.7 g. (92%) of a thicksyrup whose NMR spectrum is consistent with structure [13.3]. ##STR97##

The mesylate [13.3] (15.5 g., 0.0257 mole) was dissolved in 100 ml. ofTHF and treated with 50 ml. of 10% hydrochloric acid. After a four hourstirring period, the solution was extracted with ether and the combinedether extracts were washed with water and brine solution, dired andconcentrated on the rotary evaporator to leave 13.5 g. of an oil whoseNMR spectrum is consistent with structure [14.3].

If desired, the alcohol [14.3] can be purified by column chromatography.##STR98##

The crude alcohol [14.3] (17.4 g., 0.033 mole) dissolved in 170 ml. ofreagent grade acetone was treated, under vigorous stirring, with a totalof 12.3 ml. of Jones Reagent (CrO₃ --H₂ SO₄ --H₂ O, 0.033 mole) at sucha rate that the Reagent was consumed before the next drop was added(orange to green). The mixture was diluted with water and extracted withether. The combined ether layers were extracted with 140 ml. of ice-cold1% NaOH solution, the basic extracts acidified immediately with ice-cold10% hydrochloric acid and extracted with ether. Concentration of theneutral ether extracts gave 10.8 g. of an oil consisting mainly inunreacted [14.3]. These were reoxidized as above. The combined etherextracts containing the acidic compound were dried and concentrated togive a total of 4.6 g. of crude enol [15.3] as an oil whose IR and NMRare consistent with structure [15.3].

EXAMPLE 1 Benzyl 7β-Azido-3-acetoxymethyl-Δ³-0-2-isocephem-4-carboxylate (from dibromide intermediate) ##STR99##

The mixture of dibromide starting materials [12.1 and 13.1] (1.7 g.; 3mmole) in 50 ml. of dimethylformamide (DMF) was stirred with potassiumacetate¹ (1.2 g.; 3.5 equiv.) for 16 hours at room temperature. Themixture was then diluted with 100 ml. of diethylether and 100 ml. of 10%hydrochloric acid. The ether layer was washed with 10% sodiumbicarbonate solution (once; 50 ml.) and with brine (once; 75 ml.). Theextract was then dried and evaporated in vacuo to leave 1.0 g. of a darkbrown residue. This product was purified by passing through a shortcolumn of activity III silica gel with methylene chloride elution. Thisgave a fraction containing 700 mg. (63% yield) of a colorless oil whosespectral data confirm that it has the structure of the title product.

Crystallization from 2:1 benzene:petroleum ether gave a white solid;m.p. 94°-97° C.

Anal. Calc'd for C₁₇ H₁₆ N₄ O₆ : C, 54.84; H, 4.33; N, 15.05. Found: C,55.19; H, 4.47; N, 14.89.

EXAMPLE 2 Benzyl 7β-Azido-3-acetoxmethyl-Δ³ -O-2-isocephem-4-carboxylate (from diiodide intermediate)

Diiodide intermediate [23.1] (600 mg., 0.93 mmole) prepared according tothe method described above was added to a solution of potassium acetate(380 mg., 3.9 mmole) in 5 ml. of dimethylformamide and stirred at 24° C.for 20 hours. The solution was mixed with ether (25 ml.) and washed withwater (5 × 25 ml.). The ether mixture was dried (sodium sulfate) andevaporated in vacuo to give crude benzyl 7β-azido-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate. NMR indicated approximately 35% yield. NMRand TLC established that the product is identical with that produced inExample 1.

EXAMPLE 3 Benzyl 7β-Azido-3-formyloxymethyl-Δ³-0-2-isocephem-4-carboxylate ##STR100##

Sodium formate (2.86 g., 42 mmole) was added to a solution of thedibromide mixture [12.1 and 13.1] (2.86 g., 5.05 mmole) in DMF (100 ml.)containing water (0.1 ml.). The solution was stirred at room temperaturefor 72 hours, diluted with water (100 ml.) and extracted in ether (5 ×75 ml.). The combined organic phases were washed with brine, dried (Na₂SO₄) and concentrated to leave 2.52 g. of an oil. The oil was purifiedby column chromatography on silica gel (100 g.) eluting with 3:1 ether:petroleum ether. The main fraction contained 0.70 g. of pure benzyl7β-azido-3-formyloxymethyl-Δ³ -0-2-isocephem-4-carboxylate as acolorless oil. The NMR spectrum of the product was in agreement with thestructure of the title product.

EXAMPLE 4 ##STR101##

Dibromide mixture [12.1 and 13.1] (9.5 g., 17.2 mmole) was dissolved in100 ml. of DMF and treated with potassium phenoxyacetate (8.2 g., 43.0mmole). The reaction mixture was stirred at 50°-60° C. for 18 hours. Oncooling, it was poured onto 500 ml. of water and extracted withdichloromethane. The extracts were washed several times with water andfinally with brine. Drying over anhydrous sodium sulfate and evaporationgave crude product which was purified by "dry-column" chromatography onActivity III silica gel eluting with ether. The product was obtained asa middle cut (3.1 g., 38% yield) which was about 60% pure by NMR.

EXAMPLE 5 Benzyl 7β-Azido-3-hydroxymethyl-Δ³-0-2-isocephem-4-carboxylate ##STR102##

The crude hydroxy aldehyde starting material [14.2] (50 g., 0.117 mmole)prepared above and powdered potassium acetate (50 g., 0.51 mmole) werestirred in 350 ml. of dimethylformamide for 18 hours. The mixture wasthen diluted with water and extracted with methylene chloride. Theextract was washed with water several times and dried over anhydroussodium sulfate.

Evaporation gave 40 g. of a residue which was purified by dry-columnchromatography on silica gel (Activity III) using ether as the solvent.This gave 7,0 g. (19% yield) of pure title product as a pale-yellowsolid.

This compound was found to be identical (IR, NMR, mixed m.p.), to thecompound prepared to Example 8.

For analysis, a sample was crystallized from ethyl acetate/petroleumether (30°-60°) giving white crystals with m.p. 88°-90° C.

Anal. Calc'd. for C₁₅ H₁₄ N₄ O₅ : C, 54.54, H, 4.27; N, 16.96. Found: C,54.49; H, 4.23; N, 16.97.

EXAMPLE 6 Benzyl 7β-Azido-3-benzyloxymethyl-Δ³-0-2-isocephem-4-carboxylate ##STR103##

The crude enol starting material [15.3] (4.6 g., 0.089 mmole) wasdissolved in 75 ml. of CH₂ Cl₂, treated with triethylamine (1.55 ml.,0.11 mole) and heated under reflux for 3 hours. After cooling, themixture was washed with 10% hydrochloric acid, water, 3% NaHCO₃ solutionand brine solution. Drying (Na₂ SO₄) and concentration left 3.56 g. ofcrude material which was purified by chromatography over silica gel togive 1.05 g. (28% yield) of pure benzyl 7β-azido-3-benzyloxymethyl-Δ³-0-2-isocephem-4-carboxylate. IR and NMR of this compound were inagreement with the structure of the title product.

EXAMPLE 7 Benzyl 7β-Azido-3-acetoxymethyl-Δ³-0-2-isocephem-4-carboxylate (acetylation of benzyl7β-azido-3-hydroxymethyl-Δ³ -0-2-isocephem-4-carboxylate

Benzyl 7β-Azido-3-hydroxymethyl-Δ³ -0-2-isocephem-4-carboxylate (0.2 g.,0.6 mmole) was treated with 8 ml. of pyridine and then acetic anhydride(0.2 g., 2.0 mmole). After stirring at room temperature for 3 hours, thereaction mixture was diluted with water and extracted with ether. Theextract was washed with cold 5% hydrochloric acid, water, 1% sodiumbicarbonate, and brine. It was then dried over anhydrous sodium sulfateand evaporated to give 0.18 g. (83% yield) of the acetylated titleproduct.

This compound was found to be identical (IR, NMR, mixed m.p.) to thecompound prepared in Example 1.

EXAMPLE 8 Benzyl 7β-Azido-3-hydroxymethyl-Δ³-0-2-isocephem-4-carboxylate (acid hydrolysis method)

A. A solution of benzyl 7β-azido-3-formyloxymethyl-Δ³-0-2-isocephem-4-carboxylate (2.0 g.) in acetone (15 ml.), water (15ml.) and concentrated hydrochloric acid (2.0 ml.) was stirred at roomtemperature for 2.5 hours. The solution was extracted with CH₂ Cl₂ (3 ×25 ml.) and the combined organic extracts were washed with brine, dried(Na₂ SO₄) and concentrated to give 1.77 g. of an oil which was purifiedby column chromatography on silica gel (80 g.) to give 1.0 g. (52%yield) of pure title product identical (IR, NMR, m.p.) with a sampleprepared in Example 3 by cyclization.

B. The procedure of Part A was repeated except that the startingmaterial was benzyl 7β-azido-3-acetoxymethyl-Δ³-0-2-isocephem-4-carboxylate and the reaction time was 48 hours. Thetitle product was produced in 20% yield.

EXAMPLE 9 p-Nitrobenzyl 7β-Azido-3-methylsulfonyloxymethyl-Δ³-O-2-isocephem-4-carboxylate ##STR104##

A solution of methanesulfonyl chloride (0.50 ml., 6.5 mmole) in 10 ml.of methylene chloride was added dropwise with stirring to a solution ofp-nitrobenzyl 7β-azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate(2.41 g., 6.43 mmole), triethylamine (0.97 ml., 7.0 mmole) and 75 ml. ofmethylene chloride at -10°. After 1/2 hour at -10° and 1 hour at 24°,the solution was washed with 5% hydrochloric acid, 2% sodiumbicarbonate, and water (85 ml. each), then the solvent was evaporated invacuo to give the mesylate title product, 2.86 g. (98% yield), as ayellow foam. The NMR of the product was in agreement with the proposedstructure.

The p-nitrobenzyl 7β-azido-3-hydroxymethyl-Δ³-O-2-isocephem-4-carboxylate starting material used above may beprepared as follows:

1. The diiodide intermediate of the formula ##STR105## was prepared fromp-nitrobenzyl acetoacetate according to the procedures of Preparations 1and 1A (starting Materials) described above.

2. The diiodide intermediate (6.6 g., 9.6 mmole) was cyclized withpotassium formate (2.54 g., 30 mmole) in a solution of 100 ml. DMF and0.1 water at 0°. After stirring for 5 hours with the cooling bathremoved, the mixture was poured into 100 ml. of cold water and extractedwith methylene chloride. After washing with water containing a littleNaCl, drying and evaporation in vacuo, p-nitrobenzyl7β-azido-3-formyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate was recovered(5.3 g.) as a brown oil.

3. To a solution of 5.3 g. of the 3-formyloxymethyl intermediate in 53ml. of acetone was added 26 ml. of water and 3.2 ml. of 12M HCl. Themixture was stirred at 24° for 7 hours, then poured into 100 ml. waterand extracted with methylene chloride. The combined extracts were washedwith water containing a little sodium chloride, dried and evaporated invacuo to give 3.6 g. of a brown oil. The oil was absorbed from methylenechloride onto 18 g. of silica gel and placed on a 72 g. silica gelcolumn (grade 3, 5% ether). The column was eluted with 200 ml. of ether,then with ether/ethyl acetate 3:1. The major component (Rf 0.20) gave,on evaporation of the solvent in vacuo, a yellow solid which wasrecrystallized from acetone-ether to give the 3-hydroxymethyl startingmaterial of this example, 950 mg. (17.5% yield from the diiodide). m.p.147°-148° .

Anal. Cacl'd. for C₁₅ H₁₃ N₅ O₇ : C, 48.00; H, 3.49; N, 18.66. Found: C,48.11; H, 3.61; N, 18.81.

EXAMPLE 10 Conversion of p-Nitrobenzyl 7β-Azido-3-hydroxymethyl-Δ³-O-2-isocephem-4-carboxylate to lactone ##STR106##

An aqueous solution of 0.005N sodium carbonate (20 ml., 1.0 mmole) wasadded dropwise with stirring to a solution of p-nitrobenzyl7β-azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate (as prepared inExample 9) (0.38 g., 1.0 mmole) in 50 ml. tetrahydrofuran. The additiontook 15 minutes and the solution was stirred for an additional 5minutes, then diluted with 20 ml. brine and 20 ml. ether. The phaseswere separated and the organic phase was washed with water and brine,dried (Na₂ SO₄) and evaporated in vacuo to give 0.28 g. of a solid whichwas chromatographed (dry-column method) on 15 g. silica gel (activityIII) eluting with ether:ethyl acetate 3:1. The lactone (0.16 g., 72%)was obtained as a white crystalline solid. M.p. 178°-179° d (ethylacetate-ether).

Anal. Calc'd. for C₈ H₆ N₄ O₄ : C, 43.25; H, 2.72; N, 25.22. Found: C,43.10; H, 2.75; N, 25.45.

EXAMPLE 11 p-Nitrobenzyl 7β-Azido-3-carbamoyloxymethyl-Δ³-O-2-isocephem-4-carboxylate ##STR107##

p-Nitrobenzyl 7β-Azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate(375 mg., 1.0 mmoles) was dissolved in 15 ml. of benzene and treatedwith sodium cyanate (130 mg., 2.0 mmoles) followed by trifluoroaceticacid (0.16 ml., 2.1 mmoles). After stirring for 2 hours, the reactionmixture was treated with 25 ml. of water and 15 ml. of ethyl acetate.The aqueous layer was extracted once with 15 ml. of ethyl acetate andthe combined organic extracts were washed with water and brine. Dryingwas over anhydrous sodium sulfate and evaporation in vacuo gave 400 mg.(96%) of crude carbamate product which showed only a minor impurity bythin-layer chromatography (silica gel, 10% ethyl acetate in ether).

EXAMPLE 12 Benzyl 7β-azido-3-bromomethyl-Δ³ -O-2-isocephem-4-carboxylate##STR108##

To a solution of benzyl 7β-azido-3-hydroxymethyl-Δ³-O-2-isocephem-4-carboxylate in benzene is added about an equimolaramount of pyridine and a slight molar excess of phosphorus tribromide.There is produced the 3-bromomethyl title product.

EXAMPLE 13 p-Nitrobenzyl 7β-Azido-3-(1-methyltetrazol-5-yloxymethyl)-Δ³-O-2-isocephem-4-carboxylate ##STR109##

(p-Nitrobenzyl 7β-azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate(0.70 g., 1.86 mmole) was dissolved in methylene chloride (50 ml.) andtriethylamine (0.24 ml., 1.86 mmole) was added. While stirring thesolution, there was added dropwise (in 5 minutes) a solution ofmethanesulfonyl chloride (0.145 ml., 1.86 mmole) in methylene chloride(10 ml.). The resulting solution of p-nitrobenzyl7β-azido-3-methylsulfonyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate wasstirred at room temperature for 1 hour, (protected from moisture by acalcium chloride drying tube). At the end of this time, an additionalquantity of triethylamine (0.24 ml., 1.86 mmole) was added followed by asolution of 1-methyltetrazol-5-ol (0.186 g., 1.86 mmole) in methylenechloride (25 ml.). The solution was stirred at room temperature for 5days and then the methylene chloride was removed by evaporation in vacuoand replaced by chloroform (100 ml.). The solution was refluxed 20hours, then cooled, washed successively with 10% HCl, water and brine,dried (Na₂ SO₄) and evaporated to dryness leaving a dark semi-solid(0.80 g.). This was chromatographed (dry-column) on silica gel III (25g.), eluting with ether:ethyl acetate 3:1. The pure compound wasobtained as a white solid, m.p. 174°-176° d (ethyl acetate). NMR showedthe compound to be the title product.

Anal. Calc'd. for C₁₇ H₁₅ N₉ O₇ : C, 44.64; H, 3.30; N, 27.56. Found: C,44.87; H, 3.48; N, 27.59.

EXAMPLE 14 Benzyl 7β-Amino-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate ##STR110##

H₂ S gas was slowly introduced into a stirring solution of benzyl7β-azido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylate (793 mg., 2.1mmole), triethylamine (210 mg., 2.1 mmole) and 15 ml. of methylenechloride for 10 minutes. The flask was then flushed with nitrogen toremove excess H₂ S and then evaporated to dryness in vacuo. The residuewas partitioned between diethylether and 10% hydrochloric acid threetimes. The combined acid extracts were backwashed twice withdiethylether and then carefully alkalized with solid sodium bicarbonate,saturated with sodium chloride and extracted with methylene chloride(three times, 75 ml. portions). The combined extracts were washed withbrine (once, 75 ml. portion), dried (anhydrous sodium sulfate) andevaporated in vacuo to give 415 mg. (60% yield) of title product as acolorless oil. Spectral data were consistent with the assignedstructure.

Anal. Calc'd. for C₁₇ H₁₈ N₂ O₆ : C, 58.95, H, 5.24; N, 8.09. Found: C,58.39; H, 5.32; N, 7.95.

EXAMPLE 15 Benzyl 7β-Amino-3-benzyloxymethyl-Δ³-O-2-isocephem-4-carboxylate ##STR111##

Benzyl 7β-azido-3-benzyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate (1.45g., 3.5 mmole) was dissolved in 150 ml. of dry CH₂ Cl₂ and cooled to 0°C. Triethylamine (1 ml., 7 mmole) was added, and, while stirring andcooling, H₂ S gas was passed through the solution until it wassaturated. The solution was then allowed to come to room temperature in2 hours and concentrated on the rotary evaporator. The residue waspartitioned between 10% hydrochloric acid and ether. The mixture ofsolid and aqueous phase was separated and combined with the subsequentaqueous washings of the ether phase. The solids and combined aqueousphases were made basic with solid NaHCO₃ and extracted with CH₂ Cl₂. Theextracts were dried (Na₂ SO₄) and concentrated to leave 0.80 g. of crudetitle product. The NMR of the sample was in agreement with the structureproposed.

EXAMPLE 16 Benzyl 7β-Amino-3-hydroxymethyl-Δ³-O-2-isocephem-4carboxylate ##STR112##

Benzyl 7β-azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate (1.0 g.,3 mmole) was dissolved in 60 ml. of dry CH₂ Cl₂ and cooled to 0° C.Triethylamine (0.75 ml., 6 mmole) was added and, while stirring andcooling, H₂ S gas was passed through the solution until it wassaturated. The solution was stirred at room temperature for 30 minutesand concentrated. Fresh CH₂ Cl₂ (˜30 ml.) was added and the solution wasagain concentrated. This operation was repeated a second time. Theresidue was a yellow oil consisting of crude title product.

EXAMPLE 17 7β-Amino-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid##STR113##

Benzyl 7β-azido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylate (5.0 g.,13.4 mmole) was dissolved in 300 ml. of absolute ethanol and treatedwith palladium chloride (1.2 g., 7.1 mmole). It was then hydrogenated ona Parr Hydrogenator for one hour at room temperature and an initialpressure of 50 p.s.i.g. It was then filtered, washing with warm ethanol(150 ml.). The filtrate was evaporated in vacuo to yield 4.2 g. of aresidue in the form of a solidified foam. The NMR was consistent withthe hydrochloride salt of the title product.

The free amino acid title product is obtained by dissolving thehydrochloride salt in the minimum amount of water and adjusting the pHto 3.5 with cold concentrated ammonium hydroxide. The product iscollected by filtration and washed with cold water and acetone.

The amino acid title product has m.p. 275° (decomposition) and IR andNMR consistent with the desired structure.

A sample of the above compound (called BC-L66)was found to inhibit D.pneumoniae A9585 at a concentration of 8 mcg./ml., St. pyogenes A9604 at8 mcg./ml. and S. aureus A9537 at 32 mcg./ml.

EXAMPLE 18 p-Nitrobenzyl 7β-amino-3-(1-methyltetrazol-5-yloxymethyl)-Δ³-O-2-isocephem-4-carboxylate

p-Nitrobenzyl 7β-azido-3-(1-methyltetrazol-5-yloxymethyl)-Δ³-O-2-isocephem-4-carboxylate (0.26 g., 0.57 mmole) was dissolved inmethylene chloride (50 ml.) and nitrogen was bubbled through thesolution for 5 minutes. Triethylamine (0.145 ml., 1.14 mmole) was addedand the solution was stirred while hydrogen sulfide was gently bubbledinto it for about 11/2 minutes. The resulting solution was stirred atroom temperature for 11/2 hours by which time there was no more gasevolution. The excess hydrogen sulfide was removed by a stream ofnitrogen bubbled through the solution which was in turn extracted with10% HCl (10 ml.) and water (2 × 10 ml.). The combined aqueous extractswere carefully alkalized with 5% sodium bicarbonate solution, and thenextracted with methylene chloride (3 × 25 ml.). The organic extractswere washed with brine, dried (Na₂ SO₄), and evaporated on the aspiratorto give the title product as a white amorphous solid (0.15 g.).

EXAMPLE 19

Replacement of the benzyl 7β-azido-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate in the procedure of Example 14 with anequimolar weight of benzyl 7β-azido-3-formyloxymethyl-Δ³-O-2-isocephem-4-carboxylate, p-nitrobenzyl7β-azido-3-methylsulfonyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate,p-nitrobenzyl 7β-azido-3-carbamoyloxymethyl-Δ³-O-2-isocephem-4-carboxylate and benzyl 7β-azido-3-bromomethyl-Δ³-O-2-isocephem-4-carboxylate produces the corresponding 7β-amino benzylor p-nitrobenzyl esters of the above-mentioned compounds.

EXAMPLE 20

Replacement of the benzyl 7β-azido-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate in Example 17 with an equimolar weight ofbenzyl 7β-azido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate, benzyl7β-azido-3-formyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate,p-nitrobenzyl 7β-azido-3-methylsulfonyloxymethyl-Δ³-O-2-isocephem-4-carboxylate, p-nitrobenzyl7β-azido-3-carbamoyloxymethyl-Δ ³ -O-2-isocephem-4-carboxylate, benzyl7β-azido-3-bromomethyl-Δ³ -O-2-isocephem-4-carboxylate or p-nitrobenzyl7β-azido-3-(1-methyltetrazol-5-yloxymethyl)-Δ³-O-2-isocephem-4-carboxylate produces the corresponding 7βamino4-carboxylic acid compounds of the above-mentioned azido esters.

EXAMPLE 21 Pivaloyloxymethyl 7β-amino-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate

The title compound is produced according to the method of Example 2 ofU.K. Patent specification No. 1,229,453 by replacing the7-aminocephalosporanic acid used therein by 7β-amino-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylic acid.

The respective acetoxymethyl, methoxymethyl, acetonyl and phenacylesters of 7β-amino-3-acetoxymethyl-Δ³ -O-2-isocephen-4-carboxylic acidare prepared by substituting in the method above for the chloromethylpivalate used therein an equimolar weight of chloromethyl acetate,chloromethyl methyl ether, chloroacetone and phenacyl bromide,respectively.

EXAMPLE 22

The procedure of Example 21 is repeated using7β-amino-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-formyloxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-methylsulfonyloxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-carbamoyloxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino3-(1-methyltetrazol-5-yloxymethyl)-Δ³-O-2-isocephem-4-carboxylic acid or 7β-amino-3-bromomethyl-Δ³-O-2-isocephem-4-carboxylic acid in place of the7β-amino3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid usedtherein. There are produced the corresponding pivaloyloxymethyl esters.Replacement of chloromethyl pivalate by chloromethyl acetate,chloromethyl methyl ether, chloroacetone and phenacyl bromide,respectively, produces the corresponding acetoxymethyl, methoxymethyl,acetonyl and phenacyl esters.

EXAMPLE 23 Benzyl 7β-phenoxyacetamido-3-acetoxymethyl-Δ³-O-2-isocephen-4-carboxylate ##STR114##

A mixture of benzyl 7β-amino-3-acetoxymethyl-Δ³-O-2-isocephem-4-carboxylate (415 mg., 1.2 mmole) phenoxyacetic acid(183 mg., 1.2 mmole) EEDQ (325 mg., 1.3 mmole) and 15 ml. of methylenechloride¹ was stirred at room temperature for 1.5 hours. The mixture wasthen washed with 10% HCl (once, 20 ml. ), brine (once, 20 ml.), 10%NaHCO₃ solution (once, 30 ml.), brine (once, 30 ml.), and finally dried(anhydrous Na₂ SO₄) and evaporated to dryness leaving a colorless oil.This oil was triturated with diethylether to give 410 mg. of crude amidetitle product in 71% yield. This material was dry column chromatographedover activity III silica gel by eluting with diethylether to give 400mg. of title product as a white crystalline solid; m.p. 146° C. (corr.).

Anal. Calc'd for C₂₅ H₂₄ N₂ O₈ : C, 62.49; H, 5.04; N, 5.83. Found: C,62.58, H, 5.07; N, 5.83.

EXAMPLE 24 7β-Phenoxyacetamido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid ##STR115##

A mixture of benzyl 7β-phenoxyacetamido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylate (100 mg., 0.208 mmole), 100 mg. of 10% Pd-Cand 50 ml. of THF was hydrogentated for 15 minutes in a Parrhydrogenator with an initial pressure of 40 p.s.i.g. The catalyst wasfiltered off and the filtrate evaporated to dryness leaving 50 mg. ofwhite crystalline solid title product; m.p. 160°-170° C. (corr.)(decomposition) (recrystallized from ethanol.

Anal.Calc'd. for C₁₈ H₁₈ N₂ O₈ : C, 55.38; H, 4.65; N, 7.18. Found: C,55.32; H, 4.88; N, 7.13.

A sample of the title product (called BC-L53) after solution in waterand dilution with Nutrient Broth was found to exhibit the followingMinimum Inhibitory Concentrations (M.I.C.) in mcg./ml. versus theindicated microorganisms as determined by overnight incubation at 37° C.by tube dilution. Cephalexin was included as a comparison compound.

                  Table 1                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism           BC-L53     lexin                                           ______________________________________                                        D. pneumoniae                                                                              A9585     .06        .13                                         +5% serum*                                                                    Str. pyogenes                                                                              A9604     .13        .13                                         +5% serum*                                                                    S. aureus Smith**                                                                          A9537     1          .25                                         S. aureus Smith**                                                                          A9537     4          .5                                          +50% serum                                                                    S. aureus BX1633-2                                                                         A9606     1          1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                         A9606     125        4                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                           A15097    16         32                                          resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis**                                                                         A9531     2          2                                           E. coli Juhl**                                                                             A15119    16         4                                           E. coli**    A9675     125        8                                           K. pneumoniae**                                                                            A9977     16         2                                           K. pneumoniae**                                                                            A15130    >125       8                                           Pr. mirabilis**                                                                            A9900     4          4                                           Pr. morganii**                                                                             A15153    >125       >125                                        Ps. aeruginosa**                                                                           A9843A    >125       >125                                        Ser. marcescens**                                                                          A20019    >125       >125                                        Ent. cloacae A9656     >125       >125                                        Ent. cloacae A9657     63         2                                           Ent. cloacae A9659     >125       >125                                        ______________________________________                                          *50% Nutrient Broth - 45% Antibiotic Assay Broth                             **at 10.sup.-4 dilution.                                                 

EXAMPLE 25 7β-Phenoxyacetamido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid ##STR116## A. Acylation to prepare benzyl7β-phenoxyacetamido-3-benzyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylate

To benzyl 7β-amino-3-benzyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylate(0.4 g., 1.01 mmole) dissolved in 100 ml. of CH₂ Cl₂ was added EEDQ(0.25 g., 1.01 mmole) and phenoxyacetic acid (0.154 g., 1.01 mmole). Thesolution was stirred at room temperature for 2 hours, washedsuccessively with a 1% NaHCO₃ solution, 10% hydrochloric acid and brinesolution, dried over Na₂ SO₄ and concentrated on the rotary evaporatorto give 0.53 g. of crude benzyl 7β-phenoxyacetamido-3-benzyloxymethyl-Δ³-O- 2-isocephem-4-carboxylate. Purification was achieved throughchromatography over silica gel and gave 0.35 g. (66%) of pure benzylester.

B. Simultaneous de-blocking at 3- and 4-positions

The benzyl ester intermediate of Step A (0.35 g., 0.66 mmole) washydrogenated at atmospheric pressure in a total of 55 ml. of ethylacetate in the presence of 20% Pd(OH)₂ (0.35 g.). When hydrogenation wascomplete (18 hours), the catalyst was removed by filtration over Celiteand the filtrates were concentrated on the rotary evaporator to give0.18 g. of semi-solid which crystallized after trituration with ether.Recrystallization from acetone-ether gave 0.101 g. of white solid; m.p.157°-159° C. Spectral data, IR and NMR, were fully consistent with thestructure of the title product.

This compound was found to be identical (IR, NMR, mixed m.p.) to thecompound prepared in Example 26.

Anal. Calc'd. for C₁₆ H₁₆ N₂ O₇ : C, 55.17; H, 4.63; N, 8.04. Found: C,55.06; H, 4.59; N, 7.99.

A sample of the title product (called BC-L43) aster solution in waterand dilution with Nutrient Broth was found to exhibit the followingMinimum Inhibitory Concentrations (M.I.C.) in mcg./ml. versus theindicated microorganisms as determined by overnight incubation at 37° C.by tube dilution.

                  Table 2                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism           BC-L43     lexin                                           ______________________________________                                        D. pheumoniae                                                                              A9585     <.25       .13                                         +5% serum*                                                                    Str. pyogenes                                                                              A9604     <.25       .13                                         +5% serum*                                                                    S. aureus Smith**                                                                          A9537     <.25       .25                                         S. aureus Smith**                                                                          A9537     2          1                                           +50% serum                                                                    S. aureus BX1633-2                                                                         A9606     4          1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                         A9606     63         4                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                           A15097    32         63                                          resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis**                                                                         A9531     16         2                                           E. coli Juhl**                                                                             A15119 <125                                                                             8                                                      E. coli**    A9675     >125       32                                          K. pneumoniae**                                                                            A9977     63         4                                           K. pneumoniae**                                                                            A15130    >125       16                                          Pr. mirabilis**                                                                            A9900     63         4                                           Pr. morganii**                                                                             A15153    >125       >125                                        Ps. aeruginosa**                                                                           A9843A    >125       >125                                        Ser. marcescens**                                                                          A20019    >125       >125                                        Ent. cloacae A9656     >125       >125                                        Ent. cloacae A9657     >125       4                                           Ent. cloacae A9659     >125       >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              **at 10.sup.-4 dilution.                                                 

EXAMPLE 26 7β-Phenoxyacetamido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid

A. To the crude 7β-amino intermediate from Example 16 dissolved in 60ml. of CH₂ Cl₂ was added EEDQ (0.741 g., 3 mmole) and phenoxyacetic acid(0.456 g., 3 mmole). The solution was stirred at room temperature for21/2 hours, washed successively with a 5% hydrochloric acid solution,water, a 1% sodium bicarbonate solution and water, dried (Na₂ SO₄) andconcentrated to give 1.48 g. of crude benzyl7β-phenoxyacetamido-3-hydroxymethyl-Δ³ -O- 2 -isocephem-4-carboxylate.Crystallization in ether containing a trace of ethyl acetate afforded1.07 g. of the pure benzyl ester as a white solid.

B. The benzyl ester from Part A (0.25 g.) was hydrogenated in aParr-shaker in ethyl acetate (50 ml.) with 20% Pd(OH)₂ as the catalystat an initial pressure of 60 psi of hydrogen. After 30 minutes, thecatalyst was removed by filtration over Celite and the filtrates wereconcentrated to give the title product (0.20 g., 98% yield) as whitecrystals, m.p. 156°-158° C.

This compound was found to be identical (IR, NMR, mixed m.p.) to thecompound prepared in Example 25.

EXAMPLE 27 Benzyl 7β-phenoxyacetamido-3-methylsulfonylmethyl-Δ³ -O-2-isocephem-4-carboxylate

A solution of methanesulfonyl chloride (0.04 ml., 0.5 mmole) inmethylene chloride (10 ml.) was added dropwise to a cooled (5°) stirredsolution of benzyl 7β-phenoxyacetamido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate (0.219 g., 0.5 mmole) and triethylamine (0.01ml., 0.5 mmole) in methylene chloride (10 ml.). The cooling bath wasremoved and the solution was stirred for 2 hours at room temperature.After cooling, the solution was washed successively with 10%hydrochloric acid, diluted ammonia solution, water and brine. Drying andconcentration left 0.26 g. of title product as a semi-solid. NMRanalysis verified the identity of the product.

EXAMPLE 28 7β-(2-Thienylacetamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid ##STR117##

The crude amino acid hydrochloride prepared in Example 17 (4.2 g., 13.4mmole) was treated with 150 ml. of water, cooled to 0° C., and treatedslowly with sodium bicarbonate (7.5 g., 90 mmole). A solution of2-thienylacetic acid chloride (4.5 g., 29 mmole) in 50 ml. of acetonewas added slowly and the mixture stirred for an additional hour at 0° C.

It was then extracted with ether (2 × 100 ml.) and the aqueous phase wasacidified with dilute hydrochloric acid. The product was extracted intomethylene chloride and the extract was dried over anhydrous sodiumsulfate. Evaporation in vacuo gave a crude residue which solidified onseeding and trituration with ether. The yield of this crude material is2.6 g. (51% yield from the azido ester).

The product was purified by crystallization from absolute ethanol giving1.8 g. (35% yield from the azido ester) of title product with m.p. 182°(decomposition).

Anal. Calc'd. for C₁₆ H₁₆ N₂ O₇ S: C, 50.52; H, 4.24; N, 7.37; S, 8.43.Found: C, 50.34; H, 4.41; N, 7.47; S, 8.45.

U.V. λ _(max) ^(EtOH) 272, ε = 8654.

The NMR and IR were consistent with the proposed structure. M.I.C. datafor the above product (called BC-L58) is shown in the following table.

                  Table 3                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                         Cepha-  Cepha-                                   Organism            BC-L58   lexin   lothin                                   ______________________________________                                        D. pneumoniae A9585     .03      .25   .06                                    +5% serum*              .016     .5    .03                                    Str. pyogenes A9604     .03      .25   .06                                    +5% serum*              .016     .13   .03                                    S. aureus Smith**                                                                           A9537     .5       1     .13                                                            .25      .5    .06                                    S. aureus Smith**                                                                           A9537     1        1     .13                                    +50% serum              1        2     .5                                     S. aureus BX1633-2                                                                          A9606     1        1     .13                                    at 10.sup.-3 dil'n      .5       1     .13                                    S. aureus BX1633-2                                                                          A9606     2        4     .25                                    at 10.sup.-2 dil'n      1        4     .25                                    S. aureus meth.-                                                                            A15097    2        16    .5                                     resist.; at 10.sup.-3   1        4     .5                                     dil'n                                                                         Sal. enteritidis**                                                                          A9531     .5       2     .25                                                            .13      4     .25                                    E. coli Juhl**                                                                              A15119    8        8     16                                                             8        8     16                                     E. coli**     A9675     63       16    63                                                             32       16    32                                     K. pneumoniae**                                                                             A9977     1        2     .5                                                             .5       4     1                                      K. pneumonia**                                                                              A15130    63       16    16                                                             32       8     16                                     Pr. mirabilis**                                                                             A9900     1        4     .5                                                             .5       4     1                                      Pr. morganii**                                                                              A15153    125      >125  >125                                                           >125     >125  >125                                   Ps. aeruginosa**                                                                            A9843A    >125     >125  >125                                                           >125     >125  >125                                   Ser. marcescens**                                                                           A20019    >125     >125  >125                                                           >125     >125  >125                                   Ent. cloacae  A9656     >125     >125  >125                                                           >125     >125  >125                                   Ent. cloacae  A9647     16       4     2                                                              8        4     4                                      Ent. cloacae  A9659     >125     >125  >125                                                           >125     >125  >125                                   ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              **at 10.sup.-4 dilution                                                  

Compound BC-L58 was resolved into its individual isomers designatedBC-L68 (+ isomer) and BC-L69 (- isomer). M.I.C. data for the isomers isprovided below.

                  Table 4                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                             BC-     BC-     Cepha-                                                                              Cepha-                                 Organism         L68     L69     lexin lothin                                 ______________________________________                                        D. pneumoniae                                                                             A9585    .016    .5    .5    .03                                  +5% serum*                                                                    Str. pyogenes                                                                             A9604    .016    .5    .13   .03                                  +5% serum*                                                                    S. aureus Smith**                                                                         A9537    .06     4     .5    .06                                  S. aureus Smith**                                                                         A9537    .25     32    .2    .5                                   +50% serum                                                                    S. aureus BX1633-2                                                                        A9606    .13     8     1     .13                                  at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                        A9606    1       16    4     .25                                  at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                          A15097   .25     8     4     .5                                   resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis**                                                                        A9531    .03     8     4     .25                                  E. coli Juhl**                                                                            A15119   4       >125  8     16                                   E. coli**   A9675    32      >125  16    32                                   K. pneumoniae**                                                                           A9977    .13     32    4     1                                    K. pneumoniae**                                                                           A15130   16      >125  8     16                                   Pr. mirabilis**                                                                           A9900    .13     16    4     1                                    Pr. morganii**                                                                            A15153   125     >125  >125  >125                                 Ps. aeruginosa**                                                                          A9843A   >125    >125  >125  >125                                 Ser. marcescens**                                                                         A20019   >125    >125  >125  >125                                 Ent. cloacae                                                                              A9656    >125    >125  >125  >125                                 Ent. cloacae                                                                              A9657    16      >125  4     4                                    Ent. cloacae                                                                              A9659    >125    >125  >125  >125                                 ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              **at 10.sup.-4 dilution.                                                 

EXAMPLE 29 Benzyl 7β-phenoxyacetamido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylate (acylation of corresponding 3-hydroxymethylcompound)

A mixture of benzyl 7β-phenoxyacetamido-3-hydroxy-methyl-Δ³ -O-2-isocephem-4-carboxylate (1.07 g, 2.44 mmole), acetic anhydride (1 g.,9.8 mmole) and pyridine (15 ml.) was stirred at room temperature for 1.5hours. The reaction mixture was diluted with water and extracted withCH₂ Cl₂. The organic phase was washed with water, 10% HCl and water,dried and concentrated to leave a solid. Recrystallization of the solidin ethyl acetate-ether gave 1.0 g. of title product as white crystals,m.p. 144°-146°. This compound was found to be identical (mixed m.p., IRand NMR) to the product of Example 23.

The benzyl ester may be converted to the corresponding free acidcompound by the procedure of Example 24.

EXAMPLE 30

Separation of Diastereomers of ##STR118##

A solution of benzyl 7β-amino-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylate (1.86 g., 5.4 mmole),D-carbobenzoxyphenylglycine (1.54 g., 5.4 mmole) and EEDQ (1.46 g., 6.0mmole) in 50 ml. of methylene chloride was stirred at 24° for 16 hours.The resulting solution was washed with 5% sodium bicarbonate, 10%hydrochloric acid and dilute sodium chloride solutions, then dried(sodium sulfate) and evaporated in vacuo to give a yellow tar, 3.27 g.The tar was dissolved in methylene chloride and absorbed onto 16.5 g. ofsilica gel (grade III) which was placed on a chromatographic column of165 g. of silica gel. Elution with ether gave (after some front runningimpurities) the major component as a partially separated mixture of twodiastereomers. The last fraction (2.0 g.) (shown by NMR to be a 34:66ratio of isomers) was re-absorbed onto 10 g. of silica gel andchromatographed on 100 g. of silica gel. Elution with ether:hexane 9:1changing to ether gave further separation of the isomers. The lastfraction (371 mg.) was found to be a 15:85 mixture of two isomers. Themixture had α_(D) ²⁵° = +26.5° (CHCl₃). ##STR119##

To a solution of 325 mg. (0.53 mmole) of the mixture of isomers fromPart A in 8 ml. of ethyl acetate was added 8 ml. of ethanol, 0.53 ml. of1.00 M hydrochloric acid and 320 mg. of 30%palladium-on-diatomaceous-earth. The mixture was hydrogenated at 24° and1 atmosphere until uptake of hydrogen ceased. The catalyst was removedby filtration and the solvent evaporated in vacuo to give the amino acidhydrochloride salt as an amorphous solid, 210 mg. NMR showed the productto be a 15:85 ratio mixture of isomers (the 85% isomer is arbitrarilydesignated isomer (B) and to contain about 1/3 mole of ethanol and 1 to2 mole of water. The chemical purity of the product was estimated fromU.V. spectrum to be about 70%. The product had α_(D) ²⁵° = + 72.0°(DMSO) and U.V. λ _(max) ^(EtOH) 270 mm. (ε = 6600).

IR and NMR spectra were in agreement with the proposed structure.

A sample of the product (called BC-L65) was found to exhibit thefollowing Minimum Inhibitory Concentrations.

                  Table 5                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism           BC-L65     lexin                                           ______________________________________                                        D. pneumoniae                                                                              A9585     .06        .25                                         +5% serum*                                                                    Str. pyogenes                                                                              A9604     .06        .25                                         +5% serum*                                                                    S. aureus Smith**                                                                          A9537     1          1                                           S. aureus Smith**                                                                          A9537     4          2                                           +50% serum                                                                    S. aureus BX1633-2                                                                         A9606     2          4                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                         A9606     8          4                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                           A15097    4          8                                           resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis**                                                                         A9531     .5         4                                           E. coli Juhl**                                                                             A15119    1          8                                           E. coli**    A9675     4          16                                          K. pneumoniae**                                                                            A9977     1          4                                           K. pneumoniae**                                                                            A15130    8          16                                          Pr. mirabilis**                                                                            A9900     .5         8                                           Pr. morganii**                                                                             A15153    32         >125                                        Ps. auroginosa**                                                                           A9843A    >125       >125                                        Ser. marcescens**                                                                          A20019    125        >125                                        Ent. cloacae A9656     125        >125                                        Ent. cloacae A9657     2          4                                           Ent. cloacae A9659     32         >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              **at 10.sup.-4 dilution.                                                 

EXAMPLE 31

Repeating the general N-acylation procedures of Examples 23, 25, 26, 28or 30 to react the following acylating agents with7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid (or anester or salt thereof), the following products are obtained afterremoval of any functional blocking groups.

    ______________________________________                                        Acylating Agent                                                                              Product                                                        ______________________________________                                        mixed anhydride of                                                                           7β-(2-Aminomethylphenyl-                                  potassium 2-(1-carbo-                                                                        acetamido)-3-acetoxymethyl-Δ.sup.3 -                     methoxypropen-2-ylamino-                                                                     0-2-isocephem-4-carboxylic                                     methyl)phenylacetate with                                                                    acid                                                           isobutyl chloroformate                                                        α-benzoylureidophenyl-                                                                 7β-(α-Benzoylureidophenyl-                          acetic acid    acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                                2,6-dimethoxybenzoyl                                                                         7β-(2,6-Dimethoxybenzamido)-                              chloride       3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    D-anhydro-o-carboxy-                                                                         7β-(D-α-Hydroxyphenylacetamido)-                    mandelic acid  3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    3 benzyl-1,2,4-oxadiazole-                                                                   7β-[N-(Phenylacetimidoyl)-                                5-one-4-acetic acid                                                                          aminoacetamido]-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                                valeric acid   7β-Valeramido-3-acetoxy-                                                 methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                                phenylacetic acid                                                                            7β-Phenylacetamido-3-acetoxy-                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                                3-thienylacetyl chloride                                                                     7β-(3-Thienylacetamido)-3-                                               acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                    α-carboxybenzyl-phenyl-                                                                7β-[α-carboxy-α-phenyl-                       acetic acid    acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                                o-hydroxyphenylacetic                                                                        7β-(o-Hydroxyphenylacetamido)-                            acid           3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    cyanoacetic acid                                                                             7β-[cyanoacetamido]-3-                                                   acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                    α-cyanopropionic acid                                                                  7β-(α-cyanopropionamido)-3-                                        acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                    2-(2H)-tetrazoleacetic                                                                       7β-[2-(2H)-tetrazolylacetamido]-                          acid           3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    3-(o-chlorophenyl)-5-                                                                        7β-[3-(o-chlorophenyl)-5-                                 methyl-4-isoxazole-                                                                          methylisoxazol-4-ylcarboxamido]-                               carboxylic acid chloride                                                                     3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    1-(1H)-tetrazolylacetyl                                                                      7β-[1-(1H)-tetrazolylacetamido]-                          chloride       3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                    ______________________________________                                    

EXAMPLE 32

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with 7β-amino-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    ______________________________________                                        Acylating Agents Product                                                      ______________________________________                                        4-nitrophenylacetyl                                                                            7β-(4-nitrophenylacetamido)-                            chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  p-fluorophenylacetyl                                                                           7β-(p-fluorophenylacetamido)-                           chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  p-acetoxyphenylacetyl                                                                          7β-(p-acetoxyphenylacetamido)-                          chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  o-chlorophenylacetyl                                                                           7β-(o-chlorophenylacetamido)-                           chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  p-aminophenylacetyl                                                                            7β-(p-aminophenylacetamido)-                            chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  p-methylphenylacetyl                                                                           7β-(p-methylphenylacetamido)-                           chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-guanidinophenylacetyl                                                                        7β-(4-guanidinophenylacetamido)-                        chloride hydrochloride                                                                         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-isopropylphenylacetyl                                                                        7β-(4-isopropylphenylacetamido)-                        chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-methylthiophenylacetyl                                                                       7β-(4-methylthiophenylacetamido)-                       chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-cyanophenylacetyl                                                                            7β-(4-cyanophenylacetamido)-                            chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-methoxyphenylacetyl                                                                          7β-(4-methoxyphenylacetamido)-                          chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  2,6-dimethoxyphenylacetyl                                                                      7β-(2,6-dimethoxyphenylacetamido)-                      chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  3-sulfamylphenylacetyl                                                                         7β-(3-sulfamylphenylacetamido)-                         chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  2-methyl-4-chlorophenyl                                                                        7β-(2-methyl-4-chlorophenyl-                            acetyl chloride  acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              sydnone-3-acetyl chloride                                                                      7β-(sydnone-3-acetamido)-3-                                              acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            sydnone-4-acetyl chloride                                                                      7β-(sydnone-4-acetamido)-                                                3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  2-furylacetyl chloride                                                                         7β-(2-furylacetamido)-                                                   3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  3-furylacetyl chloride                                                                         7β-(3-furylacetamido)-3-                                                 acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                  1,2,5-thiadiazole-3-                                                                           7β-(1,2,5-thiadiazole-3-                                acetyl chloride  acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              1-cyclohexenylacetyl                                                                           7β-(1-cyclohexenylacetamido)-                           chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  1,4-cyclohexadienyl-                                                                           7β-(1,4-cyclohexadienyl-                                acetyl chloride  acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              3-(1,4-cyclohexadien-                                                                          7β-[3-(1,4-cyclohexadien-1-yl)                          1-yl)propionyl chloride                                                                        propionamido]-3-acetoxy-                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              isothiazol-4-yl-acetic                                                                         7β-(isothiazol-4-yl-acetamido)-                         acid             3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  isothiazol-5-yl-acetic                                                                         7β-(isothiazol-5-yl-acetamido)-                         acid             3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  isothiazol-3-yl-acetic                                                                         7β-(isothiazol-3-yl-acetamido)-                         acid             3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  5-phenyl-1,3,4-thiadiazolyl-                                                                   7β-(5-phenyl-1,3,4-thiadiazol-                          2-yl-acetyl chloride                                                                           2-yl-acetamido)-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              thiazol-2-yl-acetyl                                                                            7β-(thiazol-2-yl-acetamido)-                            chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  imidazol-2-yl-acetyl                                                                           7β-(imidazol-2-yl-acetamido)-                           chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  1,2,3-triazol-4-yl-                                                                            7β-(1,2,3-triazol-4-yl-                                 acetic acid      acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              oxazol-2-yl-acetyl                                                                             7β-oxazol-2-yl-acetamido)-                              chloride         3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  4-pyridylacetyl  7β-(4-pyridylacetamido)-3-                              chloride         acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                  3-pyridylacetyl chloride                                                                       7β-(3-pyridylacetamido)-3-                                               acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                  3-phenylpropionyl chloride                                                                     7β-(3-phenylpropionamido)-                                               3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  3-(p-chlorophenyl)propionyl                                                                    7β-[3-(p-chlorophenyl)propion-                          chloride         amido)-3-acetoxymethyl-Δ.sup.3 -0-                                      2-isocephem-4-carboxylic acid                                3-(p-methoxyphenyl)propionyl                                                                   7β-[p-methoxyphenyl)propion-                            chloride         amido)-3-acetoxymethyl-Δ.sup.3 -0-                                      2-isocephem-4-carboxylic acid                                3-(p-sulfamylphenyl)-                                                                          7β-[3-(p-sulfamylphenyl)propion-                        propionyl chloride                                                                             amido]-3-acetoxymethyl-Δ.sup.3 -0-                                      2-isocephem-4-carboxylic acid                                3-(3,4-dimethoxyphenyl)-                                                                       7β-[3-(3,4-dimethoxyphenyl)-                            propionyl chloride                                                                             propionamido]-3-acetoxy-                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              3-(p-hydroxyphenyl)-                                                                           7β-[3-(p-hydroxyphenyl)propion-                         propionic acid   amido]-3-acetoxymethyl-Δ.sup.3 -0-                                      2-isocephem-4-carboxylic acid                                3-(p-nitrophenyl)-                                                                             7β-[3-(p-nitrophenyl)propion-                           propionic acid   amido]-3-acetoxymethyl-Δ.sup.3 -0-                                      2-isocephem-4-carboxylic acid                                3-(2-thienyl)propionyl                                                                         7β-[3-(2-thienyl)propionamido]-                                          3-acetoxymethylΔ.sup.3 -0-2-                                            isocephem-4-carboxylic acid                                  3-(3-thienyl)propionyl                                                                         7β-[3-(3-thienyl)propionamido]-                                          3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  cyclohexylacetic acid                                                                          7β-(cyclohexylacetamido)-                                                3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                  3-phenyl-5-methylisoxazol-                                                                     7β-(3-phenyl-5-methylisoxazol-                          4-yl-acetic acid 4-yl-acetamido)-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              o-aminomethylphenylacetic                                                                      7β-(o-aminomethylphenyl-                                acid             acetamido)-3-acetoxymethylΔ.sup.3 -                                     0-2-isocephem-4-carboxylic acid                              3-methoxy-4-furazanacetyl                                                                      7β(3-methoxy-4-furazan-                                 chloride         acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic                                                    acid                                                         ______________________________________                                    

EXAMPLE 33

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with 7β-amino-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    ______________________________________                                        Acylating Agent Product                                                       ______________________________________                                        p-nitrophenoxyacetic                                                                          7β-(p-nitrophenoxyacetamido)-                            acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   p-fluorophenoxyacetic                                                                         7β-(p-fluorophenoxyacetamido)-                           acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   o-chlorophenoxyacetic                                                                         7β-(o-chlorophenoxyacetamido)-                           acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   p-sulfamylphenoxyacetic                                                                       7β-(p-sulfamylphenoxyacetamido)-                         acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   p-methylphenoxyacetic                                                                         7β-(p-methylphenoxyacetamido)-                           acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   4-hydroxyphenoxyacetic                                                                        7β-(4-hydroxyphenoxyacetamido)-                                          3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   2,4-dichlorophenoxyacetic                                                                     7β-(2,4-dichlorophenoxy-                                 acid            acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               2,6-dimethoxyphenoxyacetic                                                                    7β-(2,6-dimethoxyphenoxy-                                acid            acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               4-cyanophenoxyacetic acid                                                                     7β-(4-cyanophenoxyacetamido)-                                            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   α-phenoxypropionic acid                                                                 7β-(α-phenoxypropionamido)-                                        3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   α-(2-chlorophenoxy)-                                                                    7β-(α-(2-chlorophenoxy)propion-                    propionic acid  amidio]-3-acetoxymethyl-Δ.sup.3 -0-                                     2-isocephem-4-carboxylic acid                                 α-(2,4-dichlorophenoxy)-                                                                7β-[α-(2,4-dichlorophenoxy)-                       n-butyric acid  n-butyramido]-3-acetoxy-                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-phenoxyphenylacetic                                                                   7β-(α-phenoxyphenylacetamido)-                     acid            3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   αphenoxybutyric acid                                                                    7β-(α-phenoxybutyramido)-                                          3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   4-trifluoromethylphenoxy-                                                                     7β-(4-trifluoromethylphenoxy-                            acetic acid     acetamido)-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               benzyloxyacetyl chloride                                                                      7β-(benzyloxyacetamido)-                                                 3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   β-napthoxyacetyl                                                                         7β-(β-naphthoxyacetamido)-                          chloride        3-acetoxymethyl-Δ.sup.3 -0-2-                                           isocephem-4-carboxylic acid                                   ______________________________________                                    

EXAMPLE 34

Following the general N-acylation methods of the preceding examples, thecompounds listed below are prepared by acylating7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid with anacylating acid of the formula ##STR120## or a functional equivalent,e.g. acid halide, thereof.

    ______________________________________                                         ##STR121##                                                                   R.sup.a              R.sup.b   R.sup.c                                        ______________________________________                                         ##STR122##          H         H                                               ##STR123##          H         H                                               ##STR124##          H         H                                               ##STR125##          H         H                                               ##STR126##          H         H                                               ##STR127##          H         H                                               ##STR128##          H         H                                              4-pyridyl            H         H                                              3-pyridyl            H         H                                               ##STR129##          H         H                                               ##STR130##          H         H                                              imidazolyl (2)       H         H                                              imidazolinyl (2)     H         H                                              thiazolyl (2)        H         H                                              thiazolinyl (2)      H         H                                              triazolyl (2)        H         H                                              1-methyl-            H         H                                              imidazolyl (2)                                                                2-thienyl            H         H                                              3-thienyl            H         H                                              n-butyl              H         H                                              isobutyl             H         H                                              2-acetamido-         H         H                                              thiazol-5-yl                                                                  2-phenyl-1,3,4-      H         H                                              thiadiazol-5-yl                                                               2-methyl-1,3,4-      H         H                                              oxadiazol-5-yl                                                                ______________________________________                                    

EXAMPLE 35

Following the general N-acylation methods of the proceeding examples,the compounds listed below are prepared by acylation of7β-amino-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid with theappropriate acylating acid of the general formula R^(a) -COOH or afunctional equivalent thereof. ##STR131##7β-(3-phenyl-5-methyl-isoxazol-4-ylcarboxamido)-3-acetoxymethyl-.DELTA.³-O- 2-isocephem-4-carboxylic acid;

7β-[3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylcarboxamido]-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid;

7β-(2,6-dichlorobenzamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(2-phenylbenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(2-aminomethylbenzamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(2-carboxybenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(cyclopentanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(1-aminocyclohexanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(cyclohexanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(1,4-cyclohexadienylcarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(4-nitrobenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(4-methylbenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(o-methoxybenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(o-bromobenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(p-ethoxybenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(o-acetamidobenzamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(p-allylbenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(2,5-dihydroxybenzamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(2-ethoxy-1-napthamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(2-methoxy-1-naphthamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(o-dimethylaminobenzamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(benzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid;

7β-(p-chlorobenzamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(2-thienylcarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(3-thienylcarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(2-furylcarboxamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(3-furylcarboxamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylicacid;

7β-(2'-chlorocyclobutanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(3'-fluorocyclopentanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-caraboxylic acid;

7β-(3'-methylcyclopentanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(3'-methoxycyclopentanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(α-naphthamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid;

7β-(β-naphthamido)-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid;

7β-(1-aminocyclopentanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(1-aminocycloheptanecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid;

7β-(1-cyclohexenecarboxamido)-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid.

EXAMPLE 36

Repeating the general N-acylation procedures of the above examples toreact trityl chloride with 7β-amino-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester or salt thereof), there isobtained after removal of any carboxyl-protecting group,7β-triphenylmethylcarboxamido-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid.

EXAMPLE 37

Following the acylation methods of the preceeding examples and inparticular those disclosed in U.S Pat. No. 3,546,219, the compoundslisted below are prepared by reacting 7β-amino-3-acetoxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester or salt thereof), with theappropriate acylating agent. ##STR132##

R^(y)

phenyl

p-acetamidophenyl

p-methoxyphenyl

p-methylphenyl

2-methoxy-5-methylphenyl

m-chlorophenyl

o-nitrophenyl

2,4-dichlorophenyl

α-naphthyl

2-phenanthryl

p-aminophenyl

2-thienyl

p-dimethylaminophenyl.

EXAMPLE 38

Following the acylation methods of the preceeding examples and inparticular those disclosed in U.K. Pat Nos. 1,296,081 and 1,294,541, thecompounds listed below are prepared by reacting7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt thereof with an acylating agent of the formula ##STR133##or a functional equivalent thereof.

    ______________________________________                                         ##STR134##                                                                   R.sup.a      X     X'       R.sup.i                                           ______________________________________                                        phenyl       O     imino    NH.sub.2                                          2-thienyl    O     imino    NH.sub.2                                          3-thienyl    O     imino    NH.sub.2                                          m-nitrophenyl                                                                              O     imino    NH.sub.2                                          m-aminophenyl                                                                              O     imino    NH.sub.2                                          p-methylphenyl                                                                             O     imino    NH.sub.2                                          p-chlorophenyl                                                                             O     imino    NH.sub.2                                          p-methoxyphenyl                                                                            O     imino    NH.sub.2                                          p-hydroxyphenyl                                                                            O     imino    NH.sub.2                                          p-dimethylamino-                                                                           O     imino    NH.sub.2                                          phenyl                                                                        3,4-dimethoxy-                                                                             O     imino    NH.sub.2                                          phenyl                                                                        m-methoxyphenyl                                                                            O     imino    NH.sub.2                                          p-acetamidophenyl                                                                          O     imino    NH.sub.2                                          m-hydroxyphenyl                                                                            O     imino    NH.sub.2                                          3,5-dichloro-4-                                                                            O     imino    NH.sub.2                                          hydroxyphenyl                                                                 3-chloro-4-  O     imino    NH.sub.2                                          hydroxyphenyl                                                                 phenyl       O     O        2-furyl                                           2-thienyl    O     O        2-furyl                                           3-thienyl    O     O        2-furyl                                           phenyl       O     O        phenyl                                            2-thienyl    O     O        phenyl                                            phenyl       O     O        2-thienyl                                         p-chlorophenyl                                                                             O     O        2-furyl                                           p-hydroxyphenyl                                                                            O     O        2-furyl                                           3-chloro-4-  O     O        2-furyl                                           hydroxyphenyl                                                                 3,5-dichloro-4-                                                                            O     O        2-furyl                                           hydroxyphenyl                                                                 m-aminophenyl                                                                              O     O        2-furyl                                           p-methylphenyl                                                                             O     O        2-furyl                                           p-dimethylamino-                                                                           O     O        2-furyl                                           phenyl                                                                        p-methoxyphenyl                                                                            O     O        2-furyl                                           m-hydroxyphenyl                                                                            O     O        2-furyl                                           p-acetamidophenyl                                                                          O     O        2-furyl                                           m-nitrophenyl                                                                              O     O        2-furyl                                           phenyl       O     O        CH.sub.3                                          2-thienyl    O     O        CH.sub.3                                          3-thienyl    O     O        CH.sub.3                                          phenyl       O     O        CH.sub.2C.sub.6 H.sub.5                           phenyl       O     O                                                                                       ##STR135##                                       phenyl       O     O                                                                                       ##STR136##                                       phenyl       O     O                                                                                       ##STR137##                                       phenyl       O     O                                                                                       ##STR138##                                       phenyl       O     O                                                                                       ##STR139##                                       phenyl       O     O                                                                                       ##STR140##                                       phenyl       O     O                                                                                       ##STR141##                                       phenyl       O     O                                                                                       ##STR142##                                       phenyl       O     O                                                                                       ##STR143##                                       phenyl       S     O        2-furyl                                           2-thienyl    S     O        2-furyl                                           3-thienyl    S     O        2-furyl                                           p-hydroxyphenyl                                                                            S     O        CH.sub.3                                          phenyl       O     imino    phenyl                                            phenyl       O     imino    2-thienyl                                         phenyl       O     imino    2-furyl                                           3-thienyl    O     imino    phenyl                                            phenyl       O     imino                                                                                   ##STR144##                                       ______________________________________                                    

EXAMPLE 39

When 7β-amino-3-acetoxymetyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt thereof is acylated according to the procedures above andin particular those disclosed in U.S. Pat. No. 3,692,779 with an acidchloride of the formula ##STR145## there are produced the compoundslisted below.

    ______________________________________                                         ##STR146##                                                                   R.sup.i                                                                       ______________________________________                                        benzyl;                                                                        ##STR147##                                                                    ##STR148##                                                                    ##STR149##                                                                   ______________________________________                                    

dichloromethyl;

n-propyl;

cyclopentyl;

cyclohexyl;

p-chlorobenzyl;

phenyl;

2-thienyl;

3-thienyl.

EXAMPLE 40

When the 7-acylamido-3-acetxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acidcompounds of Example 39 are hydrogenated as thy the process of U.S. Pat.No. 3,692,779, there are produced the compounds listed below. ##STR150##

where R^(i) is as defined in Examle 39.

EXAMPLE 41

When 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt therof is acylated according to the procedures of theabove examples (and in particular the procedures disclosed in U.S. Pat.No. 3,646,024) with an acid chloride of the formula ##STR151## there areproduced the compounds listed below.

    ______________________________________                                         ##STR152##                                                                   R.sup.a               R.sup.i                                                 ______________________________________                                        phenyl                phenyl                                                  phenyl                2-thienyl                                               phenyl                2-furyl                                                 2-thienyl             phenyl                                                  3-thienyl             2-furyl                                                 p-hydroxyphenyl       phenyl                                                  ______________________________________                                    

EXAMPLE 42

When 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt thereof is acylated according to the procedures above andin particular according to the methods of U.S. Pat. No. 3,778,436 withan acylating agent of the formula ##STR153## or a functional equivalentthereof, there are produced the compounds listed below.

    ______________________________________                                         ##STR154##                                                                   R.sup.a               R.sup.1                                                 ______________________________________                                        3,4-dimethoxyphenyl   ethyl                                                   p-methylphenyl        2-thienyl                                               2,4-dichlorophenyl    ethyl                                                   5-methyl-3-phenyl-    ethyl                                                   isoxazol-4-yl                                                                 2-thienyl             ethyl                                                   2-furyl               2-furyl                                                 phenyl                phenyl                                                  1,4-cyclohexadien-    methyl                                                  1-yl                                                                          ______________________________________                                    

EXAMPLE 43

When the N-acylation and de-blocking procedures of Example 30 arerepeated with the N-carbobenzoxy-D-(-)-phenylglycine used thereinreplaced by an equimolar weight of the N-t-butoxycarbonylamino acylatingacids listed below, there are produced the following compounds.

    ______________________________________                                        D-(-)-α-(3,5-dichloro-4-                                                                7β-[D-(-)-α-amino-α-(3,5-                    hydroxyphenyl)-α-(t-butoxy-                                                             dichloro-4-hydroxyphenyl)-                                    carbonylamino)acetic acid                                                                     acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               D-(-)-α-(3-chloro-4-                                                                    7β-[D-(-)-α-amino-α-(3-chloro-               hydroxyphenyl)-2-(t-butoxy-                                                                   4-hydroxyphenyl)acetamido]-3-                                 carbonylamino)acetic acid                                                                     acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             D-(-)-α-(p-hydroxyphenyl)-                                                              7β-[D-(-)-α-amino-α-(p-hydroxy-              α-(t-butoxycarbonylamino)-                                                              phenyl)acetamido]-3-acetoxy-                                  acetic acid     methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               2-(N-t-butoxycarbonyl-                                                                        7β-[α-amino-α-(2,4,6-cyclo-                  amino)-2-(2,4,6-cyclo-                                                                        heptatrien-1-yl)acetamido]-3-                                 heptatrien-1-yl)acetic                                                                        acetoxymethyl-Δ.sup.3 -0-2-isocephem-                   acid            4-carboxylic acid                                             D-2-(t-butoxycarbonyl-                                                                        7β-[D-α-amino-α-(3'-hydroxy-                 amino)-2-(3'-hydroxy-                                                                         phenyl)acetamido]-3-acetoxy-                                  phenyl)acetic acid                                                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(4-acetamido-                amino)-α-4-acetamido-                                                                   phenyl)acetamido]-3-acetoxy-                                  phenylacetic acid                                                                             methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-2-(t-butoxycarbonyl-                                                                        7β-[D-α-amino-α-(1,4-cyclo-                  amino)-2-(1,4-cyclo-                                                                          hexadienyl)acetamido]-3-                                      hexadienyl)acetic acid                                                                        acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                   D-3-(t-butoxycarbonyl-                                                                        7β-[D-3'-amino-3'-(1,4-cyclo-                            amino)-3-(1,4-cyclo-                                                                          hexadienyl)propionamido]-3-                                   hexadienyl)propionic acid                                                                     acetoxy-methyl-Δ.sup.3 -0-2-                                            isocephem-4-carboxylic acid                                   D-(-)-α-(2-thienyl)-α-                                                            7β-[D-α-amino-α-(2-thienyl)-                 (t-butoxycarbonylamino)-                                                                      acetamido]-3-acetoxymethyl-Δ.sup.3 -                    acetic acid     0-2-isocephem-4-carboxylic acid                               D-(-)-α-(3-thienyl)-α-                                                            7β-[D-α-amino-α-(3-thienyl)-                 (t-butoxycarbonylamino)-                                                                      acetamido]-3-acetoxymethyl-Δ.sup.3 -                    acetic acid     0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-hydroxy-                  amino)-α-(o-hydroxyphenyl)-                                                             Phenyl)acetamido] -3-acetoxy-                                 acetic acid     methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-nitrophenyl)-             amino)-α-(p-nitrophenyl)-                                                               acetamido]-3-acetoxymethyl-Δ.sup.3 -                    acetic acid     0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-methoxy-                  amino)-α-(p-methoxyphenyl)-                                                             phenyl)acetamido]-3-acetoxy-                                  acetic acid     methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-cyanophenyl)-             amino)-α-(p-cyanophenyl)-                                                               acetamido]-3-acetoxymethyl-Δ.sup.3 -                    acetic acid     0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-methylthio-               amino)-α-(p-methylthio-                                                                 phenyl)acetamido]-3-acetoxy-                                  phenyl)acetic acid                                                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(p-isopropyl-                amino)-α-(p-isopropyl-                                                                  phenyl)acetamido]-3-acetoxy-                                  phenyl)acetic acid                                                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-sulfamyl-                 amino)-α-(o-sulfamyl-                                                                   phenyl)acetamido]-3-acetoxy-                                  phenyl)acetic acid                                                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-aminomethyl-              amino)-α-(o-aminomethyl-                                                                phenyl)acetamido]-3-acetoxy-                                  phenyl)acetic acid                                                                            methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(o-dimethyl-                 amino)-α-(o-dimethylamino-                                                              aminophenyl)acetamido]-3-                                     phenyl)acetic acid                                                                            acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                   D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(4-chloro-2-                 amino)-α-(4-chloro-2-                                                                   thienyl)acetamido]-3-acetoxy-                                 thienyl)acetic acid                                                                           methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(cyclohexyl)-                amino)-α-(cyclohexyl)-                                                                  acetamido]-3-acetoxymethyl-Δ.sup.3 -                    acetic acid     0-2-isocephem-4-carboxylic acid                               D-α-(t-butoxycarbonyl-                                                                  7β-[D-α-amino-α-(3-trifluoro-                amino)-α-(3-trifluoro-                                                                  methylphenyl)acetamido]-3-                                    methylphenyl)acetic acid                                                                      acetoxymethyl-Δ.sup.3 -0-2-                                             isocephem-4-carboxylic acid                                   ______________________________________                                    

EXAMPLE 44

When 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt thereof is acylated according to the general procedures ofthe preceeding examples with the acylating agents listed below (suitablyprotected), there are produced the following compounds

    ______________________________________                                        α-amino-α-(1-cyclohexenyl)-                                                       7β-[α-amino-α-(1-cyclohexenyl)-              acetic acid     acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-(isothiazol-4-                                                          7β-[α-amino-α-(isothiazol-4-                 yl)acetyl chloride                                                                            yl)acetamido]-3-acetoxy-                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               thiadiazol-2-yl)acetyl                                                                        thiadiazol-2-yl)acetamido]-3-                                 chloride        acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               oxadiazol-2-yl)acetyl                                                                         oxadiazol-2-yl)acetamido]-3-                                  chloride        acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             α-amino-α-(3-methyl-1,2,5-                                                        7β-[α-amino-α-(3-methyl-1,2,5-               oxadiazol-4-yl)acetyl                                                                         oxadiazol-4-yl)acetamido]-3-                                  chloride        acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             α-amino-α-(oxazol-2-yl)-                                                          7β-[α-amino-α-(oxazol-2-yl)-                 acetyl chloride acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-(1H)-tetra-                                                             7β-[α-amino-α-(1H)-tetrazolyl-               zolylacetyl chloride                                                                          acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-4-isoxazolyl-                                                           7β-[α-amino-α-(4-isoxazolyl)-                acetyl chloride acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-(2-thiazolyl)-                                                          7β-(α-amino-α-(2-thiazolyl)-                 acetyl chloride acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-(2-furyl)-                                                              7β-[α-amino-α-(2-furyl)-                     acetyl chloride acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               α-amino-α-(1,2,5-thiadiazol-                                                      7β-[α-amino-α-(1,2,5-thiadiazol-             3-yl)acetyl chloride                                                                          3-yl)acetamido]-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-amino-α-(3-furyl)acetyl                                                         7β-[α-amino-α-(3-furyl)-                     chloride        acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               ______________________________________                                    

EXAMPLE 45

When benzyl 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylate isacylated according to the general acylation procedures above with theacylating agents listed below (suitably protected if necessary), thereare produced the following compounds after removal of any protectinggroups:

    ______________________________________                                        D-3-chloromandelic acid                                                                        7β-[D-α-hydroxy-α-(3-chloro-                                 phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-2-trifluoromethyl-                                                                           7β-[D-α-hydroxy-α-(2-trifluoro-             mandelic acid    methylphenyl)acetamido]-3-                                                    acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            D-3-nitromandelic acid                                                                         7β-[D-α-hydroxy-α-(3-nitro-                                  phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-p-hydroxymandelic acid                                                                       7β-[D-α-hydroxy-α-(p-hydroxy-                                phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-3-chloro-4-hydroxy-                                                                          7β-[D-α-hydroxy-α-(3-chloro-4-              mandelic acid    hydroxyphenyl)acetamido]-3-                                                   acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            D-3,5-dichloro-4-hydroxy-                                                                      7β-[D-α-hydroxy-α-(3,5-dichloro-            mandelic acid    4-hydroxyphenyl)acetamido]-3-                                                 acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            D-o-methylaminomandelic                                                                        7β-[D-α-hydroxy-α-(o-methyl-                acid             aminophenyl)acetamido]-3-                                                     acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            D-p-methoxymandelic acid                                                                       7β-[D-α-hydroxy-α -(p-methoxy-                               phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-m-methylthiomandelic                                                                         7β-[D-α-hydroxy-α-(m-methyl-                acid             thiophenyl)acetamido]-3-                                                      acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            D-m-iodomandelic acid                                                                          7β-[D-α-hydroxy-α-(m-iodo-                                   phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              4-isoxazoleglycolic acid                                                                       7β-[α-hydroxy-α-(4-isoxazolyl)-                              acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              4-thiazoleglycolic acid                                                                        7β-[α-hydroxy-α-(4-thiazolyl)-                               acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              4-oxazoleglycolic acid                                                                         7β-[α-hydroxy-α-(4-oxazolyl)-                                acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    isocephem-4-carboxylic acid                                  3-isothiazoleglycolic acid                                                                     7β-[α-hydroxy-α-(3-isothiazolyl)-                            acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              1,2,3-triazole-4-glycolic                                                                      7β-[α-hydroxy-α-(1,2,3-triazol-             acid             4-yl)acetamido]-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              5-isoxazoleglycolic acid                                                                       7β-[α-hydroxy-α-(5-isoxazolyl)-                              acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              1,2,4-triazole-3-glycolic                                                                      7β-[α-hydroxy-α-(1,2,4-triazol-             acid             3-yl)acetamido]-3-acetoxy-                                                    methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              2-thienylglycolic acid                                                                         7β-[α-hydroxy-α-(2-thienyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              3-thienylglycolic acid                                                                         7β-[α-hydroxy-α-(3-thienyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              1,4-cyclohexadien-1-yl-                                                                        7β-[α-hydroxy-α-(1,4-cyclo-                 glycolic acid    hexadien-1-yl)acetamido]-3-                                                   acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                            1-cyclohexenylglycolic                                                                         7β-[α-hydroxy-α-(1-cyclo-                   acid             hexenyl)acetamido]-3-acetoxy-                                                 methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              2-pyrrolylglycolic acid                                                                        7β-[α-hydroxy-α-(2-pyrrolyl)-                                acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              2-furylglycolic acid                                                                           7β-[α-hydroxy-α-(2-furyl)-                                   acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              3-pyridylglycolic acid                                                                         7β-[α-hydroxy-α-(3-pyridyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                              ______________________________________                                    

EXAMPLE 46

When benzyl 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylate isacylated according to the general acylation procedures described abovewith the acylating agents listed below (suitably protected if desired),there are produced the following compounds after removal of anyprotecting groups:

    ______________________________________                                        p-hydroxyphenylmalonic                                                                        7β- -[α-carboxy-α-(p-hydroxy-                acid            phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               3-chloro-4-hydroxyphenyl-                                                                     7β-[α-carboxy-α-(3-chloro-4-                 malonic acid    hydroxyphenyl)acetamido]-3-                                                   acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             3,5-dichloro-4-hydroxy-                                                                       7β-[α-carboxy-α-(3,5-dichloro-               phenylmalonic acid                                                                            4-hydroxyphenyl)acetamido]-3-                                                 acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             o-chlorophenylmalonic                                                                         7β-[α-carboxy-α-(o-chloro-                   acid            phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               p-nitrophenylmalonic                                                                          7β-[α-carboxy-α-(p-nitro-                    acid            phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               p-acetoxyphenylmalonic                                                                        7β-[α-carboxy-α-(p-acetoxy-                  acid            phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               p-methoxyphenylmalonic                                                                        7β-[α-carboxy-α-(p-methoxy-                                  phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               p-methylthiophenylmalonic                                                                     7β-[α-carboxy-α-(p-methyl-                   acid            thiophenyl)acetamido]-3-                                                      acetoxymethyl-Δ.sup.3 0-2-isocephem-                                    4-carboxylic acid                                             p-cyanophenylmalonic                                                                          7β-[α-carboxy-α-(p-cyanophenyl)-             acid            acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               m-isopropylphenylmalonic                                                                      7β-[α-carboxy-α-(m-isopropyl-                acid            phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               o-aminomethylphenyl-                                                                          7β-[α-carboxy-α-(o-aminomethyl-              malonic acid    phenyl)acetamido]-3-acetoxy-                                                  methyl-Δ.sup.3 0-2-isocephem-4-                                         carboxylic acid                                               o-dimethylaminophenyl-                                                                        7β-[α-carboxy-α-(o-dimethyl-                 malonic acid    aminophenyl)acetamido[-3-                                                     acetoxymethyl-Δ.sup.3 0-2-isocephem-                                    4-carboxylic acid                                             2-thienylmalonic acid                                                                         7β-[α-carboxy-α-(2-thienyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               3-thienylmalonic acid                                                                         7β-[α-carboxy-α-(3-thienyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               1,4-cyclohexadienyl-                                                                          7β-[α-carboxy-α-(1,4-cyclo-                  malonic acid    hexadien-1-yl)acetamido]-3-                                                   acetoxymethyl-Δ.sup.3 -0-2-isocephem-                                   4-carboxylic acid                                             1-cyclohexenylmalonic                                                                         7β-[α-carboxy-α-(1-cyclo-                    acid            hexenyl)acetamido]-3-acetoxy-                                                 methyl-Δ.sup.3 0-2-isocephem-4-                                         carboxylic acid                                               2-furylmalonic acid                                                                           7β-[α-carboxy-α-(2-furyl)-                                   acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               4-pyridylmalonic acid                                                                         7β-[α-carboxy-α-(4-pyridyl)-                                 acetamido]-3-acetoxymethyl-Δ.sup.3 -                                    0-2-isocephem-4-carboxylic acid                               ______________________________________                                    

EXAMPLE 47

When 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or anester or salt thereof is acylated according to the general procedures ofthe preceeding examples with an acylating agent of the formula##STR155## or a functional equivalent thereof, there are produced thecompounds listed below

    ______________________________________                                        R.sup.a               Y                                                       ______________________________________                                        phenyl                guanidino                                               2-thienyl             guanidino                                               3-thienyl             guanidino                                               1,4-cyclohexadienyl   guanidino                                               cyclohexenyl          guanidino                                               p-hydroxyphenyl       guanidino                                               3-chloro-4-hydroxyphenyl                                                                            guanidino                                               3,5-dichloro-4-hydroxyphenyl                                                                        guanidino                                               phenyl                ureido                                                  2-thienyl             ureido                                                  3-thienyl             ureido                                                  1-cyclohexenyl        ureido                                                  1,4-cyclohexadienyl   ureido                                                  p-hydroxyphenyl       ureido                                                  3,5-dichloro-4-hydroxyphenyl                                                                        ureido                                                  o-aminomethylphenyl   ureido                                                  p-methylphenyl        ureido                                                  m-chlorophenyl        ureido                                                  phenyl                thioureido                                              2-thienyl             thioureido                                              3-thienyl             thioureido                                              1-cyclohexenyl        thioureido                                              1,4-cyclohexadienyl   thioureido                                              p-hydroxyphenyl       thioureido                                              3-chloro-4-hydroxyphenyl                                                                            thioureido                                              3,5-dichloro-4-hydroxyphenyl                                                                        thioureido                                              o-aminomethylphenyl   methylthioureido                                        m-chlorophenyl        allylthioureido                                         phenyl                chloro                                                  2-thienyl             bromo                                                   3-thienyl             chloro                                                  1,4-cyclohexadienyl   methoxy                                                 1-cyclohexenyl        ethoxy                                                  phenyl                phenyl                                                  2-thienyl             methoxy                                                 3-thienyl             ethoxy                                                  p-hydroxyphenyl       iodo                                                    p-trifluoromethylphenyl                                                                             methoxy                                                 3,4-dichlorophenyl    methoxy                                                 phenyl                acetoxy                                                 2-thienyl             acetoxy                                                 2-furyl               acetoxy                                                 p-nitrophenyl         acetoxy                                                 p-methoxyphenyl       acetoxy                                                 phenyl                propionyloxy                                            phenyl                cyano                                                   phenyl                SO.sub.3 H                                              phenyl                azido                                                   phenyl                methylsulfonyl                                          phenyl                5-indanyloxycarbonyl                                    p-hydroxyphenyl       5-indanyloxycarbonyl                                    3-chloro-4-hydroxyphenyl                                                                            5-indanyloxycarbonyl                                    3,5-dichloro-4-hydroxyphenyl                                                                        5-indanyloxycarbonyl                                    ______________________________________                                    

EXAMPLE 487β-(2-Thienylacetamido)-3-(1-methyltetrazol-5-yloxymethyl)-Δ.sup.3-0-2-isocephem-4-carboxylic acid ##STR156##

A mixture consisting of p-nitrobenzyl7β-amino-3-(1-methyltetrazol-5-yloxymethyl)-Δ³ -O-2-isocephem-4-carboxylate (0.15 g., 0.35 mmole), EEDQ (0.086 g., 0.35mmole) and thienylacetic acid (0.05 g., 0.35 mmole) in methylenechloride (50 ml. ) was kept at room temperature (protected from moistureby a calcium chloride drying tube) for 16 hours. It was then washedsuccessively with 10% HCl, water, 5% sodium bicarbonate and brine, dried(Na₂ SO₄) and evaporated to dryness to give 0.18 g. of an amorphoussolid which was indentified by NMR as p-nitrobenzyl7β-(2-Thienylacetamido)-3-(1-methyltetrazol-5-yloxymetyl)-Δ.sup.3 -O-2-isocephem-4-carboxylate.

The p-nitrobenzyl ester (0.18 g., 0.032 mmole) was dissolved inethylacetate (75 ml.) and n-butanol (10 ml.) in a 500 ml. Parr bottle.To this was added 20% palladium hydroxide on Celite (0.27 g.) and 0.1Nhydro. chloric acid (3.2 ml., 0.32 mmole). The mixture was then shakenunder hydrogen (initial pressure 60 psi.) for 2 1/2 hours. The solid wasfiltered off on a Celite pad and the filtrate evaporated in vacuo downto a volume of aout 2 ml., when a yellow solid separated. Ether (50 ml.) and 1% sodium bicarbonate (15 ml.) were added and, after thoroughshaking, the aqueous phase was isolated and carefully acidified with 10%phydrochloric acid. On cooling, white crystals separated and werecollected by suction filtration, washed with cold water and dired togive the title product as a white solid (0.04 g.) m.p. 190°-192° d.

Anal. Calc'd. for C₁₆ H₁₆ N₆ O₆ : C, 45.71; H, 3.84; N, 19.99.

Found: C, 45.63; H, 3.91; N, 20.11.

U.V. ε_(max) = 7,900 (THF)

                  Table 6                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism           BC-L72     lothin                                          ______________________________________                                        D. pneumoniae                                                                              A9585     .06        .06                                         +5% serum*                                                                    Str. pyogenes                                                                              A9604     .06        .06                                         +5% serum*                                                                    S. aureus Smith.sup.+                                                                      A9537     .25        .13                                         S. aureus Smith.sup.+                                                                      A9537     1          .5                                          +50% serum                                                                    S. aureus BX1633-2                                                                         A9606     8          .25                                         at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                         A9606     125        .5                                          at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                           A15097    8          1                                           resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis.sup.+                                                                     A9531     .5         .25                                         E. coli. Juhl.sup.+                                                                        A15119    32         16                                          E. coli.sup.+                                                                              A9675     125        63                                          K. pneumoniae.sup.+                                                                        A9977     2          2                                           K. pneumoniae.sup.+                                                                        A15130    63         16                                          Pr. mirabilis.sup.+                                                                        A9900     2          1                                           Pr. morganii.sup.+                                                                         A15153    125        >125                                        Ps. aeruginosa.sup.+                                                                       A9343A    >125       >125                                        Ser. marcescens.sup. +                                                                     A20019    >125       >125                                        Ent. cloacae A9656     >125       >125                                        Ent. cloacae A9657     16         4                                           Ent. cloacae A9659     >125       >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup. -4 dilution.                                           

EXAMPLE 49 7β-(2-Thienylacetamido)-3-Carbamoyloxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid ##STR157##

p-Nitrobenzyl 7β-azido-3-carbamoyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate (400 mg., 0.96 mmoles) and triethylamine (0.25ml., 1.8 mmoles) were dissolved in 30 ml. of methylene chloride.Hydrogen sulfide was gently bubbled into the solution for 1 minute withstirring. After stirring for 30 minutes, nitrogen was bubbled through toremove most of the excess H₂ S and it was then evaporated in vacuo.

The residue, p-nitrobenzyl 7β-amino-3-carbamoyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate, was dissolved in 50 ml. of methylene chlorideand treated with thienylacetic acid (150 mg., 1.05 mmoles) and EEDQ (240mg., 1.0 mmoles). The mixture was stirred at room temperature for 18hours and then washed with water, 1% sodium bicarbonate, water, 1%hydrochloric acid, water and brine. It was then dried over anhydroussodium sulfate and evaporated in vacuo to give crude product which waspurified by dry-column chromatography on silica gel (Activity III)eluting with 25% ethyl acetate in ether. The yield of p-nitrobenzyl7β-(2-thienylacetamido)-3-carbamoyloxymethyl-Δ³ -O-2-isocephem-4-carboxylate was 150 mg. (29%). NMR and IR of the productester were consistent with the proposed structure.

To a solution of 310 mg. (0.80 mmoles) of the ester in 150 ml. of ethylacetate and 50 ml. of n-butanol was added 8 ml. of 0.1N hydrochloricacid (0.80 mmoles) and 410 mg. of 20% palladium hydroxide on carbon. Themixture was hydrogenated at room temperature on a Parr apparatus at 60psig for 3 hours.

The catalyst was removed by filtration and the solvent was evaporated invacuo. The residue was slurried with ether and extracted with 1% sodiumbicarbonate. The bicarbonate extract was acidified with dilutehydrochloric acid and extracted with ethyl acetate. It was dried overanhydrous sodium sulfate and evaporated in vacuo to give a semi-solidresidue. The residue was triturated with ether and filtered to give 25mg. of a pale-yellow solid which was about 70-80% pure by spectral dataand analysis.

U.V. λ_(max).^(THF) = 270 mm (ε= 10,351). NMR and IR analyses confirmedthat the compound produced was the title product.

                  Table 7                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                      Cepha-   Cepha-                                     Organism         BC-L74   lexin    lothin                                     ______________________________________                                        D. pneumoniae                                                                             A9585    16       1      .13                                      +5% serum*                                                                    Str. pyogenes                                                                             A9604    8        .25    .06                                      +5% serum*                                                                    S. aureus Smith.sup.+                                                                     A9537    .5       1      .13                                      S. aureus Smith.sup.+                                                                     A9537    >63      2      .5                                       +50% serum                                                                    S. aureus BX1633-2                                                                        A9606    1        4      .25                                      at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                        A9606    >125     8      .5                                       at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                          A15097   32       16     1                                        resist.; at 10.sup.-3                                                         dil'n                                                                         Sal. enteritidis.sup.+                                                                    A9531    >125     4      2.5                                      E. coli Juhl .sup.+                                                                       A15119   >125     8      16                                       E. coli.sup.+                                                                             A9675    >125     16     63                                       K. pneumoniae.sup.+                                                                       A9977    >125     8      2                                        K. pneumoniae.sup.+                                                                       A15130   >125     16     32                                       Pr. mirabilis.sup.+                                                                       A9900    >125     8      1                                        Pr. morganii.sup.+                                                                        A15153   >125     >125   >125                                     Ps. aeruginosa.sup.+                                                                      A9343A   >125     >125   >125                                     Ser. marcescens.sup.+                                                                     A20019   >125     >125   >125                                     Ent. cloacae                                                                              A9656    >125     >125   >125                                     Ent. cloacae                                                                              A9657    >125     4      8                                        Ent. cloacae                                                                              A9659    >125     >125   >125                                     ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup.-4 dilution.                                            

EXAMPLE 50

When the 7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid(or ester or salt thereof) in the procedures of Examples 23, 28-30 and31-47 is replaced by an equimolar amount of7β-amino-3-formyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid,7β-amino-3-hydroxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid,7β-amino-3-bromomethyl-β³ -O- 2-isocephem-4-carboxylic acid,7β-amino-3-methylsulfonyloxymethyl-Δ³ -O-2-isocephem-4-carboxylic acid,7β-3-carbamoyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid or7β-amino-3-benzyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid (or anester or salt thereof, any reactive functional groups other than the7-amino group being suitably protected if necessary), respectively,there are produced (after any necessary de-blocking of functionalprotecting groups) the corresponding 7β-acylamino carboxylic acids ofeach of the above-named nuclei.

EXAMPLE 51

When the benzyl 7β-phenoxyacetamido-3-hydroxymethyl-Δ³ -O-2-isocephem-4-carboxylate in the procedure of Example 27 is replaced byan equimolar weight of the benzyl esters of each of the7β-acylamino-3-hydroxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acidproducts of Example 50, there are produced the corresponding7β-acylamino-3-methylsulfonyloxymethyl-Δ³ -O- 2-isocephem-4-carboxylateesters of the respective 3-hydroxymethyl compounds.

EXAMPLE 52

When the α-amino products of Examples 30 and 43-44 or 50 are reactedwith acetone according to the procedure of U.S. Pat. No. 3,303,193,there are obtained the corresponding 0-2-isocephem derivatives of theformula ##STR158## where R^(a) is a defined in the above-mentionedexamples and Q is acetoxymethyl, formyloxymethyl, hydroxymethyl,bromomethyl, methylsulfonyloxymethyl, carbamoyloxymethyl orbenzyloxymethyl, or pharmaceutically acceptable salts thereof.

EXAMPLE 53

When the α-amino products of Examples 30, 43-44 or 50 are reacted withdicyanogen or cyanogen bromide or cyanogen chloride according to theprocedure disclosed in U.S. Pat. No. 3,796,709, the correspondingα-cyanamino products are obtained.

EXAMPLE 54

When the α-amino products of Examples 30, 43-44 or 50 are reacted with atriethylamine --SO₃ complex according to the procedure of U.S. Pat. No.3,381,001, the corresponding α-sulfoamino products are obtained.

EXAMPLE 55

When the α-amino products of Examples 30, 43-44 or 50 are reacted with1-methyl-1-nitrosobiuret or a 1-methyl-5-(lower)alkyl-1-nitrosobiuretaccording to the procedure of U.S. Pat. No. 3,483,188, there areproduced the compounds of the general formula ##STR159## where R² ishydrogen or (lower)alkyl, Q is acetoxymethyl, formyloxymethyl,bromomethyl, methylsulfonyloxymethyl, carbamoyloxymethyl,benzyloxymethyl or hydroxymethyl and R^(a) is as defined in theabove-mentioned examples.

EXAMPLE 56 Preparation of7β-[α-(2-Aminomethyl-1,4-cyclohexadienyl)acetamido]-3-methyl-.DELTA.³-O- 2-isocephem-4-carboxylic acid ##STR160## A.α-(2-Aminomethyl-1,4-cyclohexadienyl)acetic acid

A solution of 16.5 g. (0.1 mole) of o-aminomethylphenylacetic acid in1.5 l of liquid ammonia (which had been treated with 50 mg. of Li toremove a trace of moisture) was slowly diluted with 500 ml. of dryt-BuOH. To the solution was added in small portions 3.4 g. (0.5 atom) ofLi over a period of 4 hours and the mixture was stirred for 16 hours atroom temperature removing the liquid ammonia in a hood and finallyevaporated to dryness below 40° C. The residue was dissolved in 500 ml.of water and the solution was chromatographed on a column of IR-120 (H⁺,700 ml.) resin and eluted with 1% NH₄ OH solution. Ninhydrin positivefractions of the eluate were combined and evaporated to dryness. Theresidue was washed with four 50 ml. portions of hot acetone andrecrystallized from 500 ml. of ethanol-water (1:1) to give 11.2 g. (67%)of α-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid as colorlessneedles. M.p. 183° C.

IR: ν _(max) ^(nuj) 1630, 1520, 1380, 1356 cm⁻¹.

NMR: δ D₂ O + K₂ CO₃ 2.72 (4H, s, ##STR161## ), 3.01 (2H, s, CH₂ CO),3.20 (2H, s, CH₂ -N), 5.78 (2H, s, ##STR162## ).

Anal. Calcd. for C₉ H₁₃ NO₂ : C, 64.65; H, 7.84; N, 8.38.

Found: C, 64.77; H, 8.06; N, 8.44.

B. α-[2-(t-Butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid

To a stirred solution of 8.0 g. (0.048 mole) ofα-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid and 3.8 g. (0.096 mole)of NaOH in 150 ml. of water was added a solution of 10.3 g. (0.072 mole)of t-butoxycarbonylazide in 80 ml. of THF and the mixture was stirredfor 18 hours at room temperature. The THF was removed under reducedpressure and the residue solution was washed with ether (2 × 100 ml.),acidified with 6 N HCl and extracted with ether (3 × 100 ml.). Thecombined extracts were washed with water (2 × 100 ml.) and a saturatedNaCl solution (100 ml.), dried with Na₂ SO₄ and evaporated to dryness.The oily residue was triturated with n-hexane to give 10.5 g. (82%) ofcolorless powder melting at 113° C.

IR:ν _(max) ^(nuj) 3370, 1715, 1640, 1530, 1280, 1160 cm⁻¹.

NMR: δ _(ppm) ^(CDCl) 3 1.45 (9H, s, t-Bu-H), 2.73 (4H, s, ##STR163## ),3.16 (2H, s, CH₂ CO), 3.76 (2H, d, 6Hz, CH₂ N) 4.90 (1H, m, NH), 5.66(2H, s, ##STR164## ), 10.6 (1H, br-s, COOH).

Anal. Calcd. for C₁₄ H₂₁ NO₄ : C, 62.90; H, 7.92; N, 5.24. Found: C,63.13; H, 8.21; N, 5.26.

C.7β-[α-(2-t-Butoxycarbonylaminomethyl-1,4-cyclohexadienyl)acetamido]-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid and2,4-dinitrophenol in ethyl acetate is added an equimolar amount ofN,N'-dicylohexylcarbodiimide. The reaction mixture is stirred at roomtemperature for 3 hours. The separated dicylcohexylurea is filtered off.The filtrate is evaporated to dryness to give the activated ester whichis dissolved in tetrahydrofuran. To this solution is added a solution of7β-amino-3-acetoxymethyl-Δ³ -O- 2-isocephem-4-carboxylic acid andtriethylamine in approximately a 1:2 molar proportion, respectively,relative to theα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid. Themixture is stirred at room temperature for several hours andconcentrated in vacuo. The concentrate is washed with ether, acidifiedwith dilute mineral acid and extracted with ethyl acetate. The extractsare washed with water and saturated NaCl solution and dried to give thetitle product.

D.7β-[α-(2-Aminomethyl-1,4-cyclohexadienyl)acetamido]-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1,4-cyclohexadienyl)acetamido]-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for one hour. To the solution is added dry ether until aprecipitate forms. The precipitate is collected by filtration, suspendedin water and adjusted to pH6 to give the title product.

EXAMPLE 577β-[α-(2-Aminomethyl-1-cyclohexenyl)acetamido]-3-acetoxymethyl-.DELTA.³-O- 2-isocephem-4-carboxylic acid ##STR165## A.[2-(N-t-Butoxycarbonylaminomethyl)-1-cyclohexen-1-yl]-acetic acid

A solution ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]-acetic acid(1.33 g., 5 mmoles) in 3% ammonium hydroxide (10 ml.) was hydrogenatedat 40 psi with palladium or charcoal (10%, 0.2 g.). A theoretical amountof hydrogen was taken up in 3 hours. The catalyst was removed and thefiltrate was acidified to pH 2 with dil. HCl and extracted with ethylacetate (2 × 50 ml.). The combined extracts were washed with water (20ml.), dried with Na₂ SO₄ and evaporated under reduced pressure to affordan oil (1.34 g.) which solidified on standing for several days.Recrystallization from n-hexane - ethyl acetate gave 1.2 g. titleproduct as colorless prisms melting at 118°-119° C.

IR: ν _(max) ^(nujol) 3450, 1730, 1660, 1510 cm⁻¹.

NMR: δ _(ppm) ^(CDCl) 3 ppm 1.58 (9H, s, t-butyl-H), 1.50-1.90 (4H, m,--CH₂ --), 1.90 - 2.20 (4H, m, allylic methylene-H), 3.18 (2H, s, CH₂--CO), 3.78 (2H, d, 6 Hz, CH₂ --N), 5.00 (1H, br-s, NH), 8.98 (1H, br-s,COOH).

Anal. Calcd. for C₁₄ H₂₃ NO₄ : C, 62.43; H, 8.61; N, 5.20. Found: C,62.12; H, 8.77; N, 5.37.

B.7β-[α-(2-t-Butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-acetoxymethyl-Δ³-O- 2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts of[2-(N-t-butoxycarbonylaminomethyl)-1-cyclohexen-1-yl]acetic acid and2,4-dinitrophenol in ethyl acetate is added an equimolar amount ofN,N'-dicyclohexylcarbodiimide. The reaction mixture is stirred for 1hour at room temperature and the precipitated dicyclohexylurea isfiltered off. The filtrate is cooled to 5° C. and poured into a coldsolution of 7β-amino-3-acetoxymethyl-Δ³ -0-2-isocephem-4-carboxylic acidand excess triethylamine in 50% aqueous THF. The mixture is stirredovernight at room temperature and washed with ether. The aqueous layeris acidified with dilute HCl to precipitate the title product.

C. 7β-[α-Aminomethyl-1-cyclohexenyl)acetamido]-3-acetoxymethyl-.DELTA.³-0-2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-acetoxymethyl-Δ³-0-2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for 1.5 hours. The mixture is diluted with ether to separate thetrifluoroacetate salt which is dissolved in water and neutralized togive the title product.

EXAMPLE 58

When the 7-amino intermediates in the acylation procedures of Examples23, 26, 28, 30, 31-50, 56 and 57 are replaced by the correspondingpivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl and phenacylesters, respectively, and the ester group of the 7-acylamido product isnot removed, there are obtained the corresponding pivaloyloxymethyl,acetoxymethyl, methoxymethyl, acetonyl and phenacyl esters,respectively, of the 7-acylamido end products.

What is claimed is:
 1. A compound having the formula ##STR166## whereinR is an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

wherein R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups defined above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)- alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfuylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C_(3-C) ₁₂ cycloalkyl in which thesubstitutents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and n is aninteger from 1-4;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR167## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C_(1-C) ₆ alkyl or (lower)alkoxy carbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR168## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

    R.sup.a X(CH.sub.2).sub.m CO--                             (vi)

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

    R.sup.a CO--                                               (vii)

wherein R^(a) is as defined under Ii); ##STR169## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR170## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; cyano; cyanamino; azido; amino; a group obtained byreacting the amino group Y with acetone, formaldehyde, acetaldehyde,butyraldehyde, acetylacetone, methyl acetoacetate, benzaldehyde,salicylaldehyde, methyl ethyl ketone or ethyl acetoacetate; hydroxy;(lower) alkoxy; carboxy; 5-indanyloxycarbonyl; triazolyl; tetrazolyl;halogeno; formyloxy; (lower)alkanoyloxy; sulfo; or sulfoamino;##STR171## wherein R^(d), R^(e) and R^(f) which may be the same ordifferent may each represent C₁ -C₆ alkyl, phenyl or phenyl substitutedby one or more chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, cyano, (lower)alkanoyloxy,(lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy,(lower)alkylthio, carboxy, di(lower)alkylamino or sulfamyl radicals;##STR172## wherein R^(a) is as defined under (i) and in addition may behydrogen, C₁ 14 C₆ alkyl, halogen-substituted C₁ -C₆ alkyl, phenethyl,phenoxymethyl, benzyl or R^(a) --CO-- and X is oxygen or sulfur;##STR173## wherein Y is as defined under (viii) and n is an integer of1-4;

    R.sup.g CH(NH.sub.2)(CH.sub.2).sub.n CO--                  (xii)

wherein n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

wherein m is O or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

    R.sup.h CO·CO--                                   (xiii)

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingchloro, bromo, iodo, fluoro, amino, di(lower)-alkylamino, (lower)alkyl,(lower)alkoxy, nitro or (lower)-alkanoylamino; ##STR174## wherein R^(a)is as defined under (i); X is oxygen or sulfur; X' is oxygen or imino;and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo (lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenylof 2-6 carbon atoms, ##STR175## n is an integer from 0 to 3 inclusiveand each of R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro, or trifluoromethyl; ##STR176##wherein R^(a) is as defined under (i) and R^(i) is as defined under(xiv); or ##STR177## wherein R^(a) is as defined under (i) and R¹ is(lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, a monocyclicheterocyclic radical having 5 or 6 atoms exclusive of hydrogen which areC, S, N or O, no more than 2 atoms being other than C, or a substitutedmonocyclic heterocyclic radical as defined above having one or morehalo, (lower)alkyl, (lower)alkoxy or phenyl substituents, Z isesterified hydroxyl of the formula

    --OCOR.sub.2

wherein R₂ is hydrogen, amino or (lower)alkyl and R" is hydrogen or aneasily cleavable ester selected from the group consisting of benzhydryl,benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl,phenacyl, acetonyl, (lower)-alkyl, triphenylmethyl, methoxymethyl,indanyl, phthalidyl, pivaloyloxymethyl and acetoxymethyl, or apharmaceutically acceptable salt thereof.
 2. A compound having theformula ##STR178## wherein R is an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

wherein R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups defined above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)-alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstitutents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C_(1-C) ₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy oramino radicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having1 or 2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and n is aninteger from 1-4;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR179## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxy carbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR180## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

    R.sup.a X(CH.sub.2).sub.m CO--

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

    R.sup.a CO--                                               (vii)

wherein R^(a) is as defined under (i); ##STR181## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR182## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo-C₁₋ C₈ alkoxy)C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; cyano; cyanamino; azido; amino; a group obtained byreacting the amino group Y with acetone, formaldehyde, acetaldehyde,butyraldehyde, acetylacetone, methyl acetoacetate, benzaldehyde,salicylaldehyde, methyl ethyl ketone or ethyl acetoacetate; hydroxy;(lower)alkoxy; carboxy; 5-indanyloxycarbonyl; triazolyl; tetrazolyl;halogeno; formyloxy; (lower)alkanoyloxy; sulfo; or sulfoamino;##STR183## wherein R^(d), R^(e) and R^(f) which may be the same ordifferent may each represent C₁ -C₆ alkyl, phenyl or phenyl substitutedby one or more chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy,(lower)alkylthio, carboxy, di(lower)alkylamino or sulfamyl radicals;##STR184## wherein R^(a) is as defined under (i) and in addition may behydrogen, C₁ -C₆ alkyl, halogen-substituted C₁ -C₆ alkyl, phenethyl,phenoxymethyl, benzyl or R^(a) --CO-- and X is oxygen or sulfur;##STR185## wherein Y is as defined under (viii) and n is an integer of1-4;

    R.sup.g CH(NH.sub.2)(CH.sub.2).sub.n CO--                  (xii)

wherein n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

wherein m is 0 or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

    R.sup.h CO·CO--                                   (xiii)

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingchloro, bromo, iodo, fluoro, amino, di(lower)alkylamino, (lower)alkyl,(lower)alkoxy, nitro or (lower)alkanoylamino; ##STR186## wherein R^(a)is as defined under (i); X is oxygen or sulfur; X' is oxygen or imino;and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo (lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenylof 2-6 carbon atoms, ##STR187## n is an integer from 0 to 3 inclusiveand each of R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro, or trifluoromethyl; ##STR188##wherein R^(a) is as defined under (i) and R^(i) is as defined undder(xiv); or ##STR189## wherein R^(a) is as defined under (i) and R¹ is(lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, a monocyclicheterocyclic radical having 5 or 6 atoms exclusive of hydrogen which areC, S, N or O, no more than 2 atoms being other than C, or a substitutedmonocyclic heterocyclic radical as defined above having one or morehalo, (lower)alkyl, (lower)alkoxy or phenyl substituents, R⁴ is --OCHO,--OCONH₂ or --OCOCH₃ and R" is hydrogen or an easily cleavable esterselected from the group consisting of benzhydryl, benzyl, p-nitrobenzyl,p-methoxybenzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl,(lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl,pivaloyloxymethyl and acetoxymethyl, or a pharmaceutically acceptablesalt thereof.
 3. A compound of claim 2 wherein R is an acyl group of theformula

    R.sup.a C.sub.n H.sub.2n CO--

in which R^(a) is phenyl; phenyl substituted by one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy or trifluoromethyl radicals; 2-thienyl; 3thienyl;tetrazolyl; sydnone-3- or sydnone-4; furyl; isothiazolyl; thiadiazolyloptionally substituted with phenyl; oxadiazolyl optionally substitutedwith phenyl; thiazolyl; imidazolyl; triazolyl; oxazolyl; pyridyl;furazan optionally substituted at the 3-position with methoxy;4-isoxazolyl optionally substituted at the 5-position with methyl and atthe 3-position with phenyl or halophenyl; 1,4-cyclohexadienyl;1-cyclohexenyl and 1-aminocyclohexyl.
 4. A compound of claim 3 wherein nis
 1. 5. A compound of claim 2 wherein R is an acyl group of the formula##STR190## in which R^(a) is phenyl and R^(b) and R^(c) which may be thesame or different each represent hydrogen, phenyl, benzyl, phenethyl orC₁ -C₆ alkyl.
 6. A compound of claim 2 wherein R is an acyl group of theformula ##STR191## in which R^(a) is phenyl; phenyl substituted with oneor more chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 3-pyridyl or 4-pyridyl; and R^(b)and R^(c) are hydrogen.
 7. A compound of claim 2 wherein R is an acylgroup of the formula

    R.sup.a CO--

in which R^(a) is phenyl; phenyl substituted with one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, (lower)alkylthio,cyano, (lower)alkoxy, (lower)alkylamino, di(lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 2-ethoxynaphthoyl;3-phenyl-5-methylisoxazol-4-yl; 3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl; or 1-aminocyclohexyl.
 8. Acompound of claim 2 wherein R is an acyl group of the formula ##STR192##in which R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups defined above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstituents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and Y ishydrazino, guanidino, ureido; substituted ureido of the formula##STR193## in which R^(p) is hydrogen or C₁ -C₈ alkyl and R^(q) ishydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino; azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde, acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate; hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; or sulfoamino.
 9. A compound of claim 2wherein R is an acyl group of the formula ##STR194## in which R^(a) is2-thienyl; 3-thienyl; phenyl; or phenyl substituted by one or morenitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro, or trifluoromethyl radicals; X is oxygen; X' isoxygen or imino; and R^(i) is (lower)alkyl, phenyl, 2-thienyl,3-thienyl, 2-furyl or 5-nitro-2-furyl.
 10. A compound of claim 9 whereinR^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl; X' is oxygen;and R^(i) is phenyl or 2-furyl.
 11. A compound of claim 2 wherein R isan acyl group of the formula ##STR195## in which Ar' is a radical of theformula ##STR196## in which R^(m), R^(n) and R^(o) are alike ordifferent and each is hydrogen, hydroxy, (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)alkylamino, (lower)alkanoyl,(lower)alkanoyloxy or phenyl and Y is amino or a group obtained byreacting the amino group with acetaldehyde, formaldehyde or acetone;fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy;guanidino; 3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido;sulfo; sulfoamino; ureido; thioureido; (lower)alkoxy; cyano; cyanoamino;or indanyloxycarbonyl.
 12. A compound of claim 11 wherein Ar' is phenyl,p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl,o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, hydroxy or carboxy.
 13. A compoundof claim 2 wherein R is an acyl group of the formula ##STR197##
 14. Acompound of claim 2 wherein R is an acyl group of the formula ##STR198##wherein U and V are alike or different and each is hydrogen, chloro orfluoro; ##STR199##
 15. An acid of claim 2 wherein R" is hydrogen and Ris phenoxyacetyl, or a pharmaceutically acceptable salt thereof.
 16. Anacid of claim 2 wherein R" is hydrogen and R is phenylacetyl, or apharmaceutically acceptable salt thereof.
 17. An acid of claim 2 whereinR" is hydrogen and R is 2-thienylacetyl, or a pharmaceutically acceptalesalt thereof.
 18. An acid of claim 2 wherein R" is hydrogen and R is3-thienylacetyl, or a pharmaceutically acceptable salt thereof.
 19. Anacid of claim 2 wherein R" is hydrogen and R is α-aminophenylacetyl orα-hydroxyphenylacetyl, or a pharmaceutically acceptable salt thereof.20. The D-isomer of an acid of claim 19 or a pharmaceutically acceptablesalt thereof.
 21. An acid of claim 2 wherein R" is hydrogen and R isα-carboxyphenylacetyl, cyanoacetyl, α-amino-α-(p-hydroxyphenyl)acetyl,α-aminoα-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl,α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxyα-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, 60-carboxy-α-(2-thienyl)acetyl, α-carboxy-α-(3-thienyl)acetyl,α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydononeacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.
 22. The D-isomer of an acid ofclaim 2 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl) acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 23. A compound having the formula ##STR200##wherein R represent an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

wherein R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazoly, pyridyl, tetrazoly, syndnone-3or -4, imidazoly, naphthoyl, quinoxalinyl, triazolyl, isothiazolyl,thiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl, furazon,pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b) substituted arylin which the aryl groups defined above under (a) are substituted by oneor more chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alklamido, cycloalkylamino,allylamido, morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radical; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstitutents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and n is aninteger from 1-4;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

wherein i n is an integer from 2-7, the alkenyl portion of said acylgroup being straight or branched and optionally interrupted by an oxygenor sulfur atom; ##STR201## wheein R^(a) is as defined above under (i)and in addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxy ccarbonyland R^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR202## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ and R^(b) and R^(c) as defined under (iv);

    R.sup.a X(CH.sub.2).sub.m CO--                             (vi)

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

    R.sup.a CO--                                               (vii)

wherein R^(a) is as defined under (i); ##STR203## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR204## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl) ureido;3-(benzoyl)ureido; cyano; cyanamino; azido, amino; a group obtained byreacting the amino group Y with acetone, formaldehyde, acetaldehyde,butyraldehyde, acetylacetone, methyl acetoacetate, benzaldehyde,salicylaldehyde, methyl ethyl ketone or ethyl acetoacetate; hydroxy;(lower)alkoxy; carboxy; 5-indanyloxycarbonyl; triazoyly; tetrazolyl,halogeno; formyloxy; (lower)alkanoyloxy; sulfo; or sulfoamino;##STR205## wherein R^(d), R^(e) and R^(I) which may be the same ordifferent may each represent C₁ -C₆ alkyl, phenyl or phenyl substitutedby one or more chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, cyano, (lower)alkanoyloxy, (lower) alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy,(lower)alklthio, carboxy, di(lower)alkylamino or sulfamyl radicals;##STR206## wherein R^(a) is as defined under (i) and in addition may behydrogen, C₁ -C₆ alkyl, halogen-substituted C₁ -C₆ alkyl, phenethyl,phenoxymethyl, benzyl or R^(a) --CO-- and X is oxygen or sulfur;##STR207## Wheren Y is as defined under (viii) and n is an integer of1-4;

    R.sup.g CH(NH.sub.2)(CH.sub.2).sub.n CO--                  (xii)

wherein n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

wherein m is 0 or an integer fom 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

    R.sup.h CO.sup.. CO--                                      (xiii)

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituted beingchloro, bromo, iodo, fluoro, amino, di(lower)-alkylamino, (lower)alkyl,(lower)alkoxy, nitro or (lower)alkanoylamino; ##STR208## wherein R^(a)is as defined under (i); X is oxygen or sulfur; X' is oxygen or imino;and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo (lower)alkyl, dichloromethyl, trichlorometyl, (lower)alkenyl of2-6 carbon atoms, ##STR209## ##STR210## n is an integer from 0 to 3inclusive and each of R^(k) and R^(j) is hydrogen nitro,di(lower)alkylamino, (lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro, iodo, bromo, fluoro, ortrifluoromethyl; ##STR211## wherein R^(a) is as defined under (i) andR^(i) is as defined under (xiv); or ##STR212## wherein R^(a) is asdefined under (i) and R^(l) is (lower)alkyl, cycloaclkyl of 3-12 carbonatoms, phenyl a monocyclic heterocyclic radical having 5 or 6 atomsexclusive of hydrogen which are C, S, N or O, no more than 2 atoms beingother than C, or a substituted monocyclic heterocyclic radical asdefined aov having one or more halo, (lower)alkyl, (lower)alkoxy orphenyl substitutents, and R" is hydrogen or an easily cleavable esterselected from the group consisting of benzhydryl, benzyl, p-nitrobenzyl,p-methoxybenzyl, trichloroethyl, trimethylsilyl, phenacyl, acetonyl,(lower)alkyl, triphenylmethyl, methoxymethyl, indanyl, phthalidyl,pivaloyloxymethyl and acetoxymetyl, or a pharmaceutically acceptablesalt thereof.
 24. A compound of claim 23 wherein R is an acyl group ofthe formulain which R^(a) is phenyl; phenyl substituted by one or morechloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy or trifluoromethyl radicals; 2-thienyl; 3-thienyl;tetrazolyl; sydnne-3-or sydnon-4; furyl; isothiazolyl; thiadiazolyloptionally substituted with phenyl; oxadiazolyl optionally substitutedwith phenyl; thiazoyl; imidazolyl; triazolyl; oxazolyl; pyridyl; furazanoptionally substituted at the 3-position with methoxy; 4-isoxazolyloptionally substituted at the 5-position with methyl and at the3-position with phenyl or halophenyl; 1,4-cyclohexadienyl;1-cyclohexenyl and 1-aminocyclohexyl.
 25. A compound of claim 24 whereinn is
 1. 26. A compound of claim 23 wherein R is an acyl group of theformula ##STR213## in which R^(a) is phenyl and R^(b) which may be thesame or different each represent hydrogen, phenyl, benzyl, phenethyl orC₁ -C₆ alkyl.
 27. A compound of claim 23 wherein R is an acyl group ofthe formula ##STR214## in which R^(a) is phenyl; phenyl substituted withone or more chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkythio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 3-pyridyl or 4-pyridyl; and R^(b)and R^(c) are hydrogen.
 28. A compound of claim 23 wherein R is an acylgroup of the formula

    R.sup.a --CO--

in which R^(a) is phenyl; phenyl substituted with one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, (lower)alkylthio,cyano, (lower)alkoxy, (lower)alkylamino, di(lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 2-ethoxynaphthoyl;3-phenyl-5-methylisoxazol-4-yl; 3-o-cholorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl; or 1-aminocyclohexyl. 29.A compound of claim 23 wherein R is an acyl group of the formula##STR215## in which R^(a) is (a) aryl selected from the group consistingof phenyl, 2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl,tetrazolyl, sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl,triazolyl, isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl,pyrazolyl, furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl;(b) substituted aryl in which the aryl groups defined above under (a)are substituted by one or more chloro, bromo, iodo, fluoro, nitro,amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstituents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and Y ishydrazino, guanidino, ureido; substituted ureido of the formula##STR216## in which R^(p) is hydrogen or C₁ -C₈ alkyl and R^(q) ishydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino;azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate, hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; or sulfoamino.
 30. A compound of claim 23wherein R is an acyl group of the formula ##STR217## in which R^(a) is2-thienyl; 3-thienyl; phenyl; or phenyl substituted by one or morenitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro, or trifluoromethyl radicals; X is oxygen; X' isoxygen or imino; and R^(i) is (lower)alkyl, phenyl, 2-thienyl,3-thienyl, 2-furyl or 5-nitro-2-furyl.
 31. A compound of claim 30wherein R^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl; X' isoxygen; and R^(i) is phenyl or 2-furyl.
 32. A compound of claim 23wherein R is an acyl group of the formula ##STR218## in which Ar' is aradical of the formula ##STR219## in which R^(m), R^(n) and R^(o) arealike or different and each is hydrogen, hydroxy, (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)alkylamino, (lower)alkanoyl,(lower)alkanoyloxy or phenyl and Y is amino or a group obtained byreacting the amino group with acetaldehyde, formaldehyde or acetone;fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy;guanidino; 3guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido; sulfo;sulfoamino; ureido; thioureido; (lower) alkoxy; cyano, cyanamino; orindanyloxycarbonyl.
 33. A compound of claim 32 wherein Ar' is phenyl,p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl,o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, hydroxy or carboxy.
 34. A compoundof claim 23 wherein R is an acyl group of the formula ##STR220##
 35. Acompound of claim 23 wherein R is an acyl group of the formula##STR221## wherein U and V are alike or different and each is hydrogen,chloro or fluoro; ##STR222##
 36. An acid of claim 23 wherein R" ishydrogen and R is phenoxyacetyl, or a pharmaceutically acceptable saltthereof.
 37. An acid of claim 23 wherein R" is hydrogen and R isphenylacetyl, or a pharmaceutically acceptable salt thereof.
 38. An acidof claim 23 wherein R" is hydrogen and R is α-aminophenylacetyl, orα-hydroxyphenylacetyl or a pharmaceutically acceptable salt thereof. 39.The D-isomer of an acid of claim 38, or a pharmaceutically acceptablesalt thereof.
 40. The pivaloyloxymethyl, methoxymethyl, phthalidyl,indanyl or acetoxymethyl ester of an acid of claim 39, or apharmaceutically acceptable salt thereof.
 41. An acid of claim 23wherein R" is hydrogen and R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)-acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-.alpha.-(2-thienyl)acetyl,α-amino-α-(3-thienyl)acetyl, α-amino-α-(1-cyclohexenyl)acetyl,α-amino-α-(1,4-cyclohexadienyl)acetyl, α-hydroxyacetyl,α-hydroxy-α-(2-thienyl)acetyl, α-hydroxy-α-(3-thienyl)acetyl,α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)-acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,α-aminomethylphenylacetyl, 1-amino- cyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.
 42. The pivaloyloxymethyl,methoxymethyl, phthalidyl, indanyl or acetoxymethyl ester of an acid ofclaim 41, or a pharmaceutically acceptable salt thereof.
 43. TheD-isomer of an acid of claim 23 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)-acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 44. The pivaloyloxymethyl, acetoxymethyl,phthalidyl, indanyl or methoxymethyl ester of an acid of claim 43, or apharmaceutically acceptable salt thereof.
 45. An acid of claim 23wherein R" is hydrogen and R is ##STR223## or a pharmaceuticallyacceptable salt thereof.
 46. An acid of claim 23 wherein R" is hydrogenand R is α-carboxyphenylacetyl, or a pharmaceutically acceptable saltthereof.
 47. An acid of claim 23 wherein R" is hydrogen and R iso-hydroxy-phenylacetyl, or a pharmaceutically acceptable salt thereof.48. An acid of claim 23 wherein R" is hydrogen and R is cyanoacetyl, ora pharmaceutically acceptable salt thereof.
 49. An acid of claim 23wherein R" is hydrogen and R is ##STR224## or a pharmaceuticallyacceptable salt thereof.
 50. An acid of claim 23 wherein R" is hydrogenand R is CH₃ CH₂ OCO--, or a pharmaceutically acceptable salt thereof.51. The D-isomer of an acid of claim 23 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl, or a pharmaceutically acceptable saltthereof.
 52. The D-isomer of an acid of claim 23 wherein R" is hydrogenand R is α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl, or apharmaceutically acceptable salt thereof.
 53. The D-isomer of an acid ofclaim 23 wherein R" is hydrogen and R isα-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 54. The D-isomer of an acid of claim 23 whereinR" is hydrogen and R is α-amino-α-(2-thienyl)acetyl, or apharmaceutically acceptable salt thereof.
 55. The D-isomer of an acid ofclaim 23 wherein R" is hydrogen and R is α-amino-α-(3-thienyl)acetyl, ora pharmaceutically acceptable salt thereof.
 56. The D-isomer of an acidof claim 23 wherein R" is hydrogen and R is ##STR225## or apharmaceutically acceptable salt thereof.
 57. The D-isomer of an acid ofclaim 23 wherein R" is hydrogen and R is ##STR226## or apharmaceutically acceptable salt thereof.
 58. The D-isomer of an acid ofclaim 23 wherein R" is hydrogen and R is α-hydroxy-α-(2-thienyl)acetyl,or a pharmaceutically acceptable salt thereof.
 59. The D-isomer of anacid of claim 23 wherein R" is hydrogen and R isα-hydroxy-α-(3-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 60. An acid of claim 23 wherein R" is hydrogen and R isα-carboxy-α-(2-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 61. An acid of claim 23 wherein R" is hydrogen and R isα-carboxy-α-(3-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 62. An acid of claim 23 wherein R" is hydrogen and R iso-aminomethylphenylacetyl, or a pharmaceutically acceptable saltthereof.
 63. An acid of claim 23 wherein R" is hydrogen and R iso-aminomethylphenylpropionyl, or a pharmaceutically acceptable saltthereof.
 64. An acid of claim 23 wherein R" is hydrogen and R is##STR227## or a pharmaceutically acceptable salt thereof.
 65. An acid ofclaim 23 wherein R" is hydrogen and R is ##STR228## or apharmaceutically acceptable salt thereof.
 66. An acid of claim 23wherein R" is hydrogen and R is ##STR229## or a pharmaceuticallyacceptable salt thereof.
 67. An acid of claim 23 wherein R" is hydrogenand R is 2-thienyl-acetyl, or a pharmaceutically acceptable saltthereof.
 68. An acid of claim 23 wherein R" is hydrogen and R is3-thienylacetyl, or a pharmaceutically acceptable salt thereof.
 69. Anacid of claim 23 wherein R" is hydrogen and R is ##STR230## or apharmaceutically acceptable salt thereof.
 70. An acid of claim 23wherein R" is hydrogen and R is ##STR231## or a pharmaceuticallyacceptable salt thereof.
 71. An acid of claim 23 wherein R" is hydrogenand R is ##STR232## or a pharmaceutically acceptable salt thereof. 72.An acid of claim 23 wherein R" is hydrogen and R is ##STR233## or apharmaceutically acceptable salt thereof.
 73. An acid of claim 23wherein R" is hydrogen and R is

    CH.sub.3 (CH.sub.2).sub.3 CO--,

or a pharmaceutically acceptable salt thereof.
 74. An acid of claim 23wherein R" is hydrogen and R is ##STR234## or a pharmaceuticallyacceptable salt thereof.
 75. An acid of claim 23 wherein R" is hydrogenand R is ##STR235## or a pharmaceutically acceptable salt thereof. 76.An acid of claim 23 wherein R" is hydrogen and R is ##STR236## or apharmaceutically acceptable salt thereof.
 77. An acid of claim 23wherein R" is hydrogen and R is ##STR237## or a pharmaceuticallyacceptable salt thereof.
 78. An acid of claim 23 wherein R" is hydrogenand R is ##STR238## or a pharmaceutically acceptable salt thereof. 79.An acid of claim 23 wherein R" is hydrogen and R is ##STR239## or apharmaceutically acceptable salt thereof.
 80. An acid of claim 23wherein R" is hydrogen and R is ##STR240## or a pharmaceuticallyacceptable salt thereof.
 81. An acid of claim 23 wherein R" is hydrogenand R is ##STR241## or a pharmaceutically acceptable salt thereof. 82.An acid of the formula ##STR242## or a physiologically hydrolyzed esteror pharmaceutically acceptable salt of said acid or ester.